IRE-1α inhibitors

ABSTRACT

The invention provides compounds which directly inhibit IRE-1α activity in vitro, prodrugs, and pharmaceutically acceptable salts thereof. Such compounds and prodrugs are useful for treating diseases associated with the unfolded protein response and can be used as single agents or in combination therapies.

This application incorporates by reference the contents of an 883 bytetext file created on Apr. 26, 2012 and named “sequencelisting.txt,”which is the sequence listing for this application.

This application claims the benefit of and incorporates by referenceSer. No. 61/257,696 filed Nov. 3, 2009.

FIELD OF THE INVENTION

The invention relates to IRE-1α inhibitors and their therapeutic uses.

BACKGROUND OF THE INVENTION

Protein folding stress in the endoplasmic reticulum of a cell initiatesa signal transduction cascade termed the unfolded protein response orUPR. A key enzyme, inositol requiring enzyme 1 (IRE-1α), relievesprotein folding stress by enhancing molecular chaperone activity andtherefore protects cells from stress induced apoptosis. Inhibitors ofIRE-1α are useful for treating at least B cell autoimmune diseases,certain cancers, and some viral infections.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A is a schematic diagram of the experiment described in Example29.

FIG. 1B and FIG. 1C are reverse images of RT-PCR products separated on4% agarose gels, which demonstrate dose-dependent inhibition of XBP-1splicing by compound 12-4 (CN-4) in liver (FIG. 1B) and kidney (FIG.1C). See Example 29.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides IRE-1α inhibitor compounds and prodrugs andpharmaceutically acceptable salts thereof. The invention also providespharmaceutical compositions and methods of using the IRE-1α inhibitorcompounds, prodrugs, and pharmaceutically acceptable salts thereoftherapeutically to treat disorders associated with the unfolded proteinresponse. Patients who can be treated include those with B cellautoimmune diseases, certain cancers, and some viral infections.

IRE-1α Inhibitor Compounds

IRE-1α inhibitor compounds of the invention directly inhibit IRE-1α. Thecompounds are understood to act through inhibition of the RNAse activityof enzyme. In particular embodiments of the invention this activity isdetected as cleavage of a human mini-XBP-1 mRNA stem-loop substrate5′-CAGUCCGCAGGACUG-3′ (SEQ ID NO:1) by IRE-1α in vitro by 10 to 100%.Other substrates also can be used to detect cleavage. See US2007/0105123.

IRE-1α inhibitor compounds of the invention can meet either or both ofthe following criteria:

-   -   a. Some compounds of the invention inhibit IRE-1α in the in        vitro assay with an IC₅₀ of approximately 0.0005-20 μM. Some of        these compounds have an IC₅₀ in this assay of approximately 1-20        μM. Others have an IC₅₀ in this assay of approximately 0.1-1 μM.        Still others have an IC₅₀ of approximately 0.0005-0.1 μm.    -   b. Some compounds of the invention inhibit IRE-1α in an in vivo        XBP-1 splicing assay (e.g., in myeloma cells) with an EC₅₀ in        the range of approximately 0.05-80 μM. Some of these compounds        have an EC0₅₀ in this assay of approximately 10-80 μM. Others        have an EC0₅₀ in this assay of approximately 1-10 μM. Still        others have an EC0₅₀ in this assay of approximately 0.05-1 μM.

DEFINITIONS

The following terms are used in this specification.

“Halogen” includes fluorine, chlorine, bromine, and iodine.

Unless otherwise specified, the term “alkyl” as used herein means asaturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5, or 6carbon atoms (“C1-C6 alkyl”) and can be linear, branched, or acombination thereof. “C1-C6 alkyl” includes C1-C5 alkyl, C1-C4 alkyl,and C1-C3 alkyl. Examples of C1-C6 alkyls include methyl, ethyl, propyl,isopropyl, sec-butyl, tert-butyl, n-butyl, 2-butyl, pentyl, and hexyl.

“Alkoxy” as used herein means —O-alkyl groups, where “alkyl” is asdefined above, and can be linear, branched, or a combination thereof.Examples of C1-C6 alkoxys include, for example, methoxy, ethoxy,propoxy, 2-propoxy, butoxy, sec-butoxy, and tert-butoxy.

The term “perfluoroalkyl” means an alkyl group as defined above in whichall of the hydrogen atoms are replaced by fluorine atoms. The term“perfluoroalkoxy” means an alkoxy group in which the alkyl moiety is aperfluoroalkyl group as defined above.

The term “hydroxylalkyl” as used herein means an alkyl group as definedabove which is substituted with a hydroxyl group.

The term “alkoxylalkyl” means radicals of the formula C_(a)H_(2a+1)—O—(CH₂)_(b)—, in which a and b independently are 1, 2, 3, 4, 5, or 6.

A “cycloalkyl” is a saturated or partially saturated 3- to 14-membered(i.e., a 3-, 4-, 5-, 6, 7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-membered)monocyclic or polycyclic ring, such as a 5-, 6-, or 7-memberedmonocyclic ring or a 10-membered bicyclic ring, in which all of the ringmembers are carbon atoms. Examples of cycloalkyls include cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

“Aryl,” when used alone or as part of another term, means a carbocyclicaromatic ring containing 5 to 14 members (e.g., 5, 6, 7, 8, 9, 10, 11,12, 13, or 14 members) and can be monocyclic or polycyclic. Examples ofaryls include phenyl, naphthyl, anthryl, and phenanthryl.

A “heterocycle,” “heterocyclic group,” and “heterocyclic ring” is asaturated or a partially saturated 4- to 14-membered (i.e., 4-, 5-, 6-,7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-membered) monocyclic or polycyclic(fused) ring, such as a 5-, 6-, or 7-membered monocyclic ring or a10-membered bicyclic ring which has 1, 2, 3, or 4 heteroatoms selectedfrom nitrogen (N), oxygen (O), and sulfur (S). Any of the nitrogen andsulfur heteroatoms optionally can be oxidized, and any nitrogenheteroatom optionally can be quaternized. A heterocyclic ring can beattached at any suitable heteroatom or carbon atom. Examples ofheterocycles include azepinyl, furyl, thienyl, pyrrolyl, pyrazolyl,imidazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazolyl,isobenzofuranyl, furazanyl, indolyl, quinolinyl, oxazolyl, imidazolinyl,isoxazolyl, quinolyl, naphthyridinyl, phenoxazinyl, phenanthridinyl,chromenyl, triazinyl, purinyl, benzothienyl, benzimidazolyl,benzopyranyl, benzothiazolyl, benzoazolyl, benzo[b]thienyl,naphtho[2,3-b]-thienyl, isothiazolyl, thiazolyl, isothiazolyl,isoquinolinyl, thiadiazolyl, oxadiazolyl, tetrahydroquinolinyl,indolizinyl, isoindolyl, indazolyl, isoquinolyl, phthalazinyl,tetrahydroquinolinyl, and cinnolinyl.

A “heteroaryl” is a saturated 4- to 14-membered (i.e., 4-, 5-, 6-, 7-,8-, 9-, 10-, 11-, 12-, 13-, or 14-membered) monocyclic or polycyclic(fused) ring, such as a 5-, 6-, or 7-membered monocyclic ring or a10-membered bicyclic ring which has 1, 2, 3, or 4 heteroatoms selectedfrom nitrogen (N), oxygen (O), and sulfur (S). Any of the nitrogen andsulfur heteroatoms optionally can be oxidized, and any nitrogenheteroatom optionally can be quaternized. A heteroaryl can be attachedat any suitable heteroatom or carbon atom. Examples of heteroarylsinclude pyridyl, imidazolyl, pyrrolyl, thienyl, furyl, pyranyl,pyrimidinyl, pyridazinyl, indolyl, quinolyl, naphthyridinyl, andisoxazolyl.

Compounds

Compounds of the invention fall into one or more of the structuralformulae described below. Non-limiting examples of compounds fallingwithin the scope of these formulae are provided in Table 1 and in theExamples.

Some embodiments of the invention include only compounds which havestructural formula (1):

which encompasses formula (1a), (1b), (1c), and (1d), in which:

in formula (1a):

-   -   R3, R4, and R8 independently are hydrogen, halogen,        perfluoroalkyl, —CN, —CONH₂, —CON(CH₃)₂, alkyl, perfluoroalkoxy,        alkoxy, hydroxylalkyl, or alkoxylalkyl;    -   R5 is hydrogen or R7;    -   R6 is hydrogen, halogen, perfluoroalkyl, perfluoroalkoxy, —CN,        alkyl, alkoxy, hydroxylalkyl, or alkoxylalkyl;    -   R7 is halogen; —CN; —CONH₂; —CON(CH₃)₂; alkyl; perfluoroalkyl;        alkoxy; hydroxylalkyl; alkoxylalkyl; perfluoroalkoxyl;

-   -    phenyl, optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl, alkoxylalkyl,        perfluoroalkoxyl,

-   -    a 5- or 6-membered heteroaryl that is substituted with 1, 2, or        3 substituents independently selected from the group consisting        of halogen, —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl,        alkoxylalkyl, perfluoroalkoxyl,

-   -    amino,

-   -    wherein n is 0, 1, or 2;

-   -   R9 is alkyl; alkoxylalkyl; perfluoroalkoxylalkyl; aryl,        optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5- or 6-membered        heterocycle, optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5- or 6-membered        heteroaryl, optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; or

wherein n is 0, 1, 2, or 3; and R10 is hydrogen or R9;or

-   -   R9 and R10, together with the nitrogen atom to which they are        attached, form a heterocycle containing 1, 2, 3, or 4        heteroatoms selected from N, O and S, optionally substituted        with 1, 2, or 3 substituents selected independently from R11;    -   R11 is hydrogen; alkyl; aryl; heteroaryl containing 1 or 2        heteroatoms selected from N, O, and S; arylalkyl;        heteroarylalkyl in which the heteroaryl contains 1 or 2        heteroatoms selected from N, O, and S;

-   -   R12 is amino; alkoxy; aryl, optionally substituted with 1, 2, or        3 substitutents selected independently from R11; a 5- or        6-membered heterocycle having 1, 2, or 3 heteroatoms selected        from N, O, and S and optionally substituted with 1, 2, or 3        substitutents selected independently from R11; or a 5- or        6-membered heteroaryl having 1, 2, or 3 heteroatoms selected        from N, O, and S and optionally substituted with 1, 2, or 3        substitutents selected independently from R11;    -   R13 is alkyl; alkoxylalkyl; perfluoroalkoxylalkyl; aryl,        optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxyl, —CN, —CONH₂, —CON(CH₃)₂,        alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5- or        6-membered heterocycle, optionally substituted with 1, 2, or 3        substituents independently selected from the group consisting of        halogen, perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂,        —CON(CH₃)₂, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5-        or 6-membered heteroaryl, optionally substituted with 1, 2, or 3        substituents independently selected from the group consisting of        halogen, perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂,        —CON(CH₃)₂, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl; or

wherein n is 0, 1, 2, or 3; and R14 is hydrogen or R13;or

-   -   R13 and R14, together with the nitrogen to which they are        attached, form a heterocycle containing 1, 2, or 3 heteroatoms        selected independently from N, O, and S, optionally substituted        with 1, 2, or 3 substitutents selected independently from R16;    -   R15 is amino; alkoxy; aryl, optionally substituted with 1, 2, or        3 substitutents selected independently from the group consisting        of halogen, perfluoroalkyl, perfluoroalkoxyl, —CN, —CONH₂,        —CON(CH₃)₂, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5-        or 6-membered heterocycle having 1, 2, or 3 heteroatoms selected        from N, O, and S and optionally substituted with 1, 2, or 3        substitutents selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; or a 5- or 6-membered        heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and        S and optionally substituted with 1, 2, or 3 substitutents        selected from the group consisting of halogen, perfluoroalkyl,        perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl, alkoxy,        hydroxylalkyl, and alkoxylalkyl;    -   R16 is hydrogen; alkyl; aryl; heteroaryl containing 1 or 2        heteroatoms selected from N, O, and S; arylalkyl;        heteroarylalkyl in which the heteroaryl contains 1 or 2        heteroatoms selected from N, O, and S;

-   -    amino; or

-   -   R17 is alkyl; alkoxylalkyl; perfluoroalkoxylalkyl; aryl,        optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5- or 6-membered        heterocycle, optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        —CN, —CONH₂, —CON(CH₃)₂, perfluoroalkyl, perfluoroalkoxy, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5- or 6-membered        heteroaryl, optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        perfluoroalkyl, perfluoroalkoxy, —CN, —CONH₂, —CON(CH₃)₂, alkyl,        alkoxy, hydroxylalkyl, and alkoxylalkyl; or

wherein n is 0, 1, 2, or 3; and R18 is hydrogen or R17;or

-   -   R17 and R18, together with the nitrogen to which they are        attached, form a heterocycle containing 1, 2, 3, or 4        heteroatoms selected independently from N, O, and S, optionally        substituted with 1, 2, or 3 substituents selected independently        from R20;    -   R19 is alkoxy; aryl, optionally substituted with 1, 2, or 3        substitutents selected independently from the group consisting        of halogen, perfluoroalkyl, perfluoroalkoxyl, —CN, —CONH₂,        —CON(CH₃)₂, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl; a 5-        or 6-membered heterocycle, optionally substituted with 1, 2, or        3 substituents independently selected from the group consisting        of halogen, —CN, —CONH₂, —CON(CH₃)₂, perfluoroalkyl,        perfluoroalkoxy, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl;        or a 5- or 6-membered heteroaryl, optionally substituted with 1,        2, or 3 substituents independently selected from the group        consisting of halogen, perfluoroalkyl, perfluoroalkoxy, —CN,        —CONH₂, —CON(CH₃)₂, alkyl, alkoxy, hydroxylalkyl, and        alkoxylalkyl;    -   R20 is halogen; perfluoroalkyl; perfluoroalkoxy; —CN; —CONH₂;        —CON(CH₃)₂; alkyl; alkoxy; hydroxylalkyl; alkoxylalkyl; and a 5-        or 6-membered heterocycle having 1 or 2 heteroatoms selected        from N, O, and S and optionally substituted with 1, 2, or 3        substituents selected independently from the group consisting of        halogen, —CN, —CONH₂, —CON(CH₃)₂, perfluoroalkyl,        perfluoroalkoxy, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl;        or a 5- or 6-membered heteroaryl, optionally substituted with 1,        2, or 3 substituents selected independently from the group        consisting of halogen, —CN, —CONH₂, —CON(CH₃)₂, perfluoroalkyl,        perfluoroalkoxy, alkyl, alkoxy, hydroxylalkyl, and alkoxylalkyl,    -   with the exception of compounds in which R5, R6, R7, and R8 are        independently hydrogen, halogen, —CH₃, —OCH₃, or hydroxymethyl;

in formula (1b):

-   -   R3, R4, R5, and R8 are hydrogen;    -   R6 and R7 independently are

-   -    wherein n is 0, 1, or 2;

-   -    and    -   R9 and R10 are as defined above in connection with formula (1a),        except that R7 and R6 cannot both be methoxy;

in formula (1c):

-   -   R3, R4, and R8 independently are hydrogen, halogen, —CN, —CONH₂,        —CON(CH₃)₂, alkyl, C2-C6 alkoxy, hydroxylalkyl, or alkoxylalkyl;    -   R5, R6, and R7 independently are hydrogen; halogen; —CN; —CONH₂;        —CON(CH₃)₂; alkyl; perfluoroalkyl; C2-C6 alkoxy; hydroxylalkyl;        alkoxylalkyl; perfluoroalkoxyl;

-   -    phenyl, optionally substituted with 1, 2, or 3 substituents        independently selected from the group consisting of halogen,        —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl, alkoxylalkyl,        perfluoroalkoxyl,

-   -    a 5- or 6-membered heteroaryl that is optionally mono-, or di-,        or tri-substituted with halogen, —CN, alkyl, perfluoroalkyl,        alkoxy, hydroxylalkyl, alkoxylalkyl, perfluoroalkoxyl,

-   -    wherein n is 0, 1, or 2;

-   -    and    -   R9, R10, R11, and R12 are as defined above in connection with        formula (1a), provided that either: (1) at least one of R3, R4,        R5, and R8 is not hydrogen; or (2) if each of R4, R5, R6, R7,        and R8 is hydrogen, R3 is not hydrogen, methoxy, or

-   -    and

in formula (1d):

-   -   R3, R4, and R8 independently are hydrogen; halogen;        perfluoroalkyl; —CN; —CONH₂; —CON(CH₃)₂; perfluoroalkoxy; alkyl;        alkoxy; hydroxylalkyl; alkoxylalkyl;    -   R5, R6, and R7, provided that neither [R5, R6, and R7] nor [R3,        R4, R5, R7, and R8] are simultaneously hydrogen, independently        are hydrogen; halogen; —CN; —CONH₂; —CON(CH₃)₂; alkyl;        perfluoroalkyl; C2-C6 alkoxy; hydroxylalkyl; alkoxylalkyl;        perfluoroalkoxyl;

-   -    phenyl, optionally substituted with 1, 2, or 3 substitutents        independently selected from the group consisting of —CN,        perfluoroalkyl, alkoxy, alkoxylalkyl, perfluoroalkoxyl, and

-   -    a 5- or 6-membered heteroaryl that is substituted with 1, 2, or        3 substitutents independently selected from the group consisting        of —CN, alkyl, perfluoroalkyl, hydroxylalkyl, alkoxylalkyl,        perfluoroalkoxyl,

-   -    wherein n is 0, 1, or 2;

-   -    and    -   R9, R10, R11, and R12 are as defined above in connection with        formula (1a), with the proviso that if R3, R4, R5, R8 and one of        R6 and R7 are hydrogen, and the other of R6 and R7 is

-   -    then R12 is not phenyl.

Examples of

include the following, in which “X” is halogen, —CN, —CONH₂, —CON(CH₃)₂,C1-C4 alkyl, C1-C4 alkoxy, C1-C4 hydroxylalkyl, or C1-C4 alkoxylalkyl:

Another embodiment includes only those compounds of formula (1a) inwhich R6 is perfluoroalkyl, perfluoroalkoxy, —CN, alkyl, alkoxy,hydroxylalkyl, or alkoxylalkyl; and R7 is —CN; —CONH₂; —CON(CH₃)₂;alkyl; perfluoroalkyl; alkoxy; hydroxylalkyl; alkoxylalkyl;perfluoroalkoxyl; phenyl, optionally substituted with 1, 2, or 3substituents independently selected from the group consisting ofhalogen, —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl,alkoxylalkyl, perfluoroalkoxyl,

a 5- or 6-membered heteroaryl attached via a carbon atom and that issubstituted with 1, 2, or 3 substituents independently selected from thegroup consisting of halogen, —CN, alkyl, perfluoroalkyl, alkoxy,hydroxylalkyl, alkoxylalkyl, perfluoroalkoxyl,

amino,

wherein n is 0, 1, or 2; and

Another embodiment includes only those compounds of formula (1a) inwhich R5 is R7, and R7 is

Another embodiment includes only those compounds of formula (1a) inwhich R5 is R7, and R7 is

Another embodiment includes only those compounds of formula (1a) inwhich R5 is R7, and R7 is

Another embodiment includes only those compounds of formula (1a) inwhich R9 and R10, together with the nitrogen atom to which they areattached, form a fused heterocycle containing 1, 2, 3, or 4 heteroatomsselected from N, O and S, with fewer than 12 atoms total, optionallysubstituted with 1, 2, or 3 substituents independently selected fromR11.

Another embodiment includes only those compounds of formula (1a) inwhich R11 is aryl.

Another embodiment includes only those compounds of formula (1a) inwhich R11 is heteroaryl containing 1 or 2 heteroatoms selected from N,O, and S.

Another embodiment includes only those compounds of formula (1a) inwhich R11 is arylalkyl.

Another embodiment includes only those compounds of formula (1a) inwhich R11 is arylalkyl containing 1 or 2 heteroatoms selected from N, O,and S.

Another embodiment includes only those compounds of formula (1a) inwhich one or more of the alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl,alkoxylalkyl, and perfluoroalkoxyl groups are C1-C4 alkyl, C1-C4perfluoroalkyl, C1-C4 alkoxy, C1-C4 hydroxylalkyl, C1-C4 alkoxylalkyl,or C1-C4 perfluoroalkoxyl groups.

Another embodiment includes only those compounds of formula (1a) inwhich R4 and R8 are hydrogen. Among these compounds are those in whichR5 is R7, and R7 is phenyl, optionally substituted with 1, 2, or 3substituents independently selected from the group consisting ofhalogen, —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl,alkoxylalkyl, perfluoroalkoxyl,

or a 5- or 6-membered heteroaryl that is substituted with 1, 2, or 3substituents independently selected from the group consisting ofhalogen, —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl,alkoxylalkyl, perfluoroalkoxyl,

Another embodiment includes only those compounds of formula (1a) inwhich R4 and R8 are hydrogen; R5 is R7, and R7 is

wherein n is 0, 1, or 2; or

Another embodiment includes only those compounds of formula (1a) inwhich R4 and R8 are hydrogen; R5 is R7, and R7 is

Another embodiment includes only those compounds of formula (1b) inwhich R6 and R7 independently are

wherein n is 0, 1, or 2;

Another embodiment includes only those compounds of formula (1b) inwhich R6 and R7 independently are

Another embodiment includes only those compounds of formula (1b) inwhich R6 and R7 independently are

Another embodiment includes only those compounds of formula (1b) inwhich R6 and R7 independently are

Another embodiment includes only those compounds of formula (1c) inwhich R5, R6, and R7 independently are

Another embodiment includes only those compounds of formula (1c) inwhich R5, R6, and R7 independently are —CN; —CONH₂; —CON(CH₃)₂; alkyl;perfluoroalkyl; C2-C6 alkoxy; hydroxylalkyl; alkoxylalkyl;perfluoroalkoxyl; phenyl, optionally substituted with 1, 2, or 3substituents independently selected from the group consisting ofhalogen, —CN, alkyl, perfluoroalkyl, alkoxy, hydroxylalkyl,alkoxylalkyl, perfluoroalkoxyl,

a 5- or 6-membered heteroaryl attached via a carbon and optionallymono-, or di-, or tri-substituted with halogen, —CN, alkyl,perfluoroalkyl, alkoxy, hydroxylalkyl, alkoxylalkyl, perfluoroalkoxyl,

wherein n is 0, 1, or 2; or

Another embodiment includes only those compounds of formula (1c) inwhich R5, R6, and R7 independently are

Another embodiment includes only those compounds of formula (1c) inwhich R5, R6, and R7 independently are

Another embodiment includes only those compounds of formula (1c) inwhich one or more of the alkyls, alkoxys, hydroxylalkyls, oralkoxylalkyls in formula (1d) independently are C1-C4 alkyl, C1-C4alkoxy, C1-C4 hydroxylalkyl, or C1-C4 alkoxylalkyl.

Another embodiment includes only those compounds of formula (1d) inwhich R5, R6, and R7 independently are —CN; —CONH₂; —CON(CH₃)₂; alkyl;perfluoroalkyl; C2-C6 alkoxy; hydroxylalkyl; alkoxylalkyl;perfluoroalkoxyl; phenyl, optionally substituted with 1, 2, or 3substitutents independently selected from the group consisting of —CN,perfluoroalkyl, alkoxy, alkoxylalkyl, perfluoroalkoxyl, and

a 5- or 6-membered heteroaryl that is substituted with 1, 2, or 3substitutents independently selected from the group consisting of —CN,alkyl, perfluoroalkyl, hydroxylalkyl, alkoxylalkyl, perfluoroalkoxyl,

wherein n is 0, 1, or 2;

Another embodiment includes only those compounds of formula (1d) inwhich R5, R6, and R7 independently are

Another embodiment includes only those compounds of formula (1d) inwhich R5, R6, and R7 independently are

Another embodiment includes only those compounds of formula (1d) inwhich R5, R6, and R7 independently are

Another embodiment includes only those compounds of formula (1d) inwhich one or more of the alkyls, alkoxys, hydroxylalkyls, oralkoxylalkyls independently are C1-C4 alkyl, C2-C6 or C2-C4 alkoxy,C1-C4 hydroxylalkyl, or C1-C4 alkoxylalkyl.

Some embodiments of the invention include only compounds which havestructural formula (2):

which encompasses structural formulae (2a), (2b), and (2c), wherein:in formula (2a):

-   -   R3 is hydrogen; halogen; or alkyl;    -   R6 is

-   -   A is        -   (a) a 4-, 5-, or 6-membered saturated cycloalkyl; or        -   (b) a 4-, 5-, or 6-membered saturated heterocycle containing            1 or 2 heteroatoms selected from N, O, and S; and    -   R11 is as defined above in connection with formula (1a);        in formula (2b):    -   R3 is hydrogen or —CN;    -   R6 is

-   -   Het is a five-membered heteroaryl containing 1, 2, or 3        heteroatoms atoms selected from N, S, and O and optionally        substituted with alkyl, provided that Het is not unsubstituted

-   -   R24 is —OH or

-   -   R9 and R10 independently are alkyl; or    -   R9 and R10, together with the atoms to which they are attached,        form a 4-, 5-, 6-, or 7-membered heteroaryl or heterocycle        containing 1 or 2 heteroatoms selected from N, O and S,        optionally substituted with alkyl; or    -   R9 is hydrogen and R10 is

-   -    wherein n is 0, 1, 2, or 3; and    -   R25 is C1-C3 alkoxy or a 5- or 6-membered heteroaryl or        heterocycle having one or two heteroatoms selected from N, O,        and S and optionally substituted with alkyl, with the proviso        that when Het is

-   -    then R24 is not —OH,

-   -    and        in formula (2c):    -   R3 is —OH, —CN, halogen, C1-C6 alkyl,

-   -    wherein n is 0, 1, or 2;    -   R6 is hydrogen or halogen; and    -   R9 and R10 are as defined above in connection with formula (1a).

Some embodiments include only those compounds of formula (2) in which R6is linked via a carbon atom.

Some embodiments include only those compounds of formula (2) in which R6is linked via a nitrogen atom.

Some embodiments include only those compounds of formula (2a) in whichR3 is C1-C6 alkyl.

Another embodiment includes only those compounds of formula (2a) inwhich A is attached via a carbon atom.

Another embodiment includes only those compounds of formula (2a) inwhich A is a 4-, 5-, or 6-membered saturated heterocycle containing anitrogen atom and is attached via the nitrogen atom.

Another embodiment includes only those compounds of formula (2a) inwhich A is

and R11 is as defined above in connection with formula (1a). In some ofthese compounds, R11 is selected from the group consisting of hydrogen;alkyl;

aryl; heteroaryl containing 1 or 2 heteroatoms selected from N, O, andS; arylalkyl; heteroarylalkyl in which the heteroaryl contains 1 or 2heteroatoms selected from N, O, and S;

and R13 and R14 are as defined above in connection with formula (1a). Insome of these compounds, the alkyl is C1-C6 or C1-C4 alkyl; thearylalkyl is aryl-C1-C6- or aryl-C1-C4 alkyl; and/or R3 is hydrogen,halogen, or alkyl (including C1-C6 and C1-C4 alkyl).

Another embodiment includes only those compounds of formula (2a) inwhich A is

R11 is

and R13 and R14 are as defined above in connection with formula (1a). Insome compounds, the alkyl is C1-C4 alkyl. In some compounds, R3 ishydrogen.

Another embodiment includes only those compounds of formula (2a) inwhich A is

in which R13 is

wherein n is 0, 1, 2, or 3; R14 is hydrogen or methyl; and R12 is C1-C3alkoxy or a 6-membered heterocycle containing 0, 1, or 2 heteroatomsselected from N, O, and S.

Another embodiment includes only those compounds of formula (2a) inwhich A is

R23 is carbon or nitrogen; R11 is hydrogen,

and R13 and R14 are as defined above in connection with formula (1a).

Another embodiment includes only those compounds of formula (2a) inwhich R13 is methyl; benzyl; or

and R14 is R13 or hydrogen.

Another embodiment includes only those compounds of formula (2a) inwhich R13 and R14, together with the nitrogen to which they areattached, form a 6-membered heterocycle containing 1, 2, or 3heteroatoms selected from N, O, and S, optionally substituted with C1-C6or C1-C4 alkyl.

Another embodiment includes only those compounds of formula (2a) inwhich one or more of the alkyls, alkoxys, hydroxylalkyls, oralkoxylalkyls in formula (1d) independently are C1-C4 alkyl, C1-C4alkoxy, C1-C4 hydroxylalkyl, or C1-C4 alkoxylalkyl.

Another embodiment includes only those compounds of formula (2b) inwhich Het is linked via a carbon atom.

Another embodiment includes only those compounds of formula (2b) inwhich Het is linked via a nitrogen atom.

Another embodiment includes only those compounds of formula (2b) inwhich one or more of the alkyls in formula (5b) is C1-C4 alkyl.

Another embodiment includes only those compounds of formula (2c) inwhich R3 is —CN, C1-C6 alkyl,

wherein n is 0, 1, or 2.

Another embodiment includes only those compounds of formula (2c) inwhich R3 is

Another embodiment includes only those compounds of formula (2c) inwhich R3 is

Some embodiments of the invention include only compounds which havestructural formula (3):

which encompasses formula (3a), (3b), (3c), (3d), (3e), (3f), (3g), and(3h), in which R6 is selected from the group consisting of:

in which

-   -   Het is a five-membered heteroaryl containing 1, 2, or 3 atoms        selected from N, S, and O and optionally substituted with alkyl;    -   n is 0 or 1; and    -   R9 and R10, together with the nitrogen atom to which they are        attached, form a 4-, 5-, 6-, or 7-membered heterocycle        containing 1 or 2 heteroatoms selected from N, O, and S,        optionally substituted with alkyl;

-   -   wherein R9 and R10 are independently but not simultaneously        hydrogen; or independently C1-C6 linear alkyl or C6 branched        alkyl;

-   -   wherein R9 and R10 are as defined above in connection with        formula (1a); and R26 is hydrogen or —NH₂;    -   (3d) pyrimidine substituted with halogen, C1-C3 alkoxy,

-   -    R9 and R10, together with the nitrogen atom to which they are        attached, form a 4-, 5-, 6-, or 7-membered saturated heterocycle        containing 1 or 2 heteroatoms and optionally substituted with        alkyl; or R9 and R10 independently are alkyl; or R9 is hydrogen        and R10 is

-   -    wherein n is 0, 1, 2, or 3; and R12 is alkoxy or a 5- or        6-membered saturated heterocycle having one or two heteroatoms        selected from O, N, and S and optionally substituted with alkyl;

-   -    in which R27 is —OH, alkoxy, or

-   -    R9 and R10 independently are hydrogen, methyl, benzyl, or

-   -    or R9 and R10, together with the nitrogen to which they are        attached, form a 6-membered heterocycle containing 1 or 2        heteroatoms selected from N, O, and S, optionally substituted        with alkyl;

-   -    in which R30 is hydrogen or halogen; one of R28 and R29 is        hydrogen and the other is

-   -    R31 is —OH or

-   -    R9 and R10 independently are hydrogen, methyl, benzyl, or

-   -    or R9 and R10, together with the nitrogen to which they are        attached, form a 6-membered saturated heterocycle, optionally        substituted with C1-C3 alkyl, provided that either (1) R30 and        R28 are not both hydrogen; or (2) R30 and R29 are not both        hydrogen;

-   -   in which R32 is alkoxy, —OH, or

-   -    R12 is hydrogen and R11 is benzyl, optionally substituted with        C1-C3 alkoxy; cyclohexane; a 6-membered saturated heterocycle        with 1 or 2 heteroatoms selected from O, N, and S; or phenyl,        optionally substituted with 1-methyl-piperazine or        dimethyl-piperazine; or R11 and R12, together with the nitrogen        atom to which they are attached, form a six-membered heterocycle        containing 2 heteroatoms selected from N, O, and S, optionally        substituted with C1-C3 alkyl or phenyl; and n is 1, 2, or 3; and

in which R33 is C2-C6 alkyl; C2-C6 alkoxylalkyl; C2-C6perfluoroalkoxylalkyl; aryl; a 5- or 6-membered heterocycle bondingthrough a carbon; or a 5- or 6-membered heteroaryl bonding through acarbon. Any of the C2-C6 alkyl, the C2-C6 alkoxylalkyl, the C2-C6perfluoroalkoxylalkyl, the aryl, the 5- or 6-membered heterocycle, orthe 5- or 6-membered heteroaryl are optionally substituted with 1, 2, or3 substituents independently selected from the group consisting ofhalogen, perfluoroalkyl, perfluoroalkoxy, —CN, alkyl, alkoxy,hydroxylalkyl, alkoxylalkyl and

wherein n is 0, 1, or 2;

and R9 and R10 are as defined above in connection with formula (1a).

Another embodiment includes only those compounds of formula (3) in whichR6 is linked via a carbon atom.

Another embodiment includes only those compounds of formula (3) in whichR6 is linked via a nitrogen atom.

Another embodiment includes only those compounds of formula (3a) inwhich Het is linked via a carbon atom.

Another embodiment includes only those compounds of formula (3a) inwhich Het is substituted with C1-C6 or C1-C4 alkyl.

Another embodiment includes only those compounds of formula (3d) inwhich R6 is attached via a carbon atom.

Another embodiment includes only those compounds of formula (3d) inwhich R6 is attached via a nitrogen atom.

Another embodiment includes only those compounds of formula (3d) inwhich one or more of the alkyls is C1-C4 alkyl.

Another embodiment includes only those compounds of formula (3d) inwhich one or more of the alkoxys is C1-C4 alkoxy.

Another embodiment includes only those compounds of formula (3e) inwhich one or more of the alkyls is C1-C4 alkyl.

Another embodiment includes only those compounds of formula (3e) inwhich the alkoxy is C1-C4 alkoxy.

Another embodiment includes only those compounds of formula (3g) inwhich n is 2.

Another embodiment includes only those compounds of formula (3g) inwhich n is 2, R32 is —OH, alkoxy, or

-   -    and R11 and R12 are independently hydrogen, C1-C4 alkyl,        benzyl, or

-   -    or R11 and R12, together with the nitrogen to which they are        attached, form a 6-membered saturated heterocycle, optionally        substituted with C1-C6 alkyl.

Some embodiments of the invention include only compounds which havestructural formula (4):

in which

-   -   R5 is

-   -    and    -   R9 and R10 are as defined above in connection with formula (1a).

Another embodiment includes only those compounds of formula (4) in whichR9 is not —OH or methoxy.

Another embodiment includes only those compounds of formula (4) in whichR5 is

Another embodiment includes only those compounds of formula (4) in whichR5 is

Another embodiment includes only those compounds of formula (4) in whichR5 is

Table 1 provides examples of compounds encompassed by one or more of thestructural formulas described above. In Table 1, “CHO” indicates

“Bn” is benzyl; “Ph” is phenyl; and “Me” is methyl. The average IC₅₀ andEC₅₀ were determined as described in the Examples below.

TABLE 1 IC50_av EC50_av Synthesis Compound Structure (μM) (μM) Example 1

<0.1 <10 12-16 2

<1 <10 12-17 3

<1 <10 12-18 4

<0.1 <10 19-2  5

<0.1 <10 12-19 6

<0.1 >10 19-11 7

<1 10 19-22 8

<0.1 <10 19-3  9

>1 >10 19-12 10

<1 <10 19-4  11

<1 10 19-15 12

<1 10 19-18 13

<0.1 >10 19-14 14

<0.1 <10 19-23 15

<1 >10 19-16 16

<0.1 10 19-5  17

<1 >10 19-13 18

<1 10 17-2  19

<0.1 nd 17-1  20

<1 nd 17-3  21

0.1 <10 18-1  22

<1 <10 15-1  23

<1 10 15-2  24

>1 >10 19-1  25

<0.1 >10 19-17 26

>1 >10 19-6  27

<1 >10 19-7  28

>1 >10 19-8  29

<1 >10 19-9  30

<1 >10 19-10 31

<0.1 >10 15-4  32

<0.1 <10 15-3  33

<0.1 <10 19-24 34

<0.1 <10 19-19 35

<0.1 <10 19-25 36

<0.1 <10 19-26 37

<0.1 <10 19-20 38

<1 <10 19-21 39

<1 >10 16-2  40

>1 >10 16-1  41

<0.1 <10 14-2  42

<0.1 <10 14-3  43

<0.1 <10 14-4  44

>1 <10 19-44 45

<1 <10 14-5  46

<0.1 <10 14-6  47

<0.1 <10 14-7  48

<0.1 <10 14-8  49

<0.1 <10 14-9  50

<1 <10 19-31 51

<1 <10 16-4  52

<1 <10 16-3  53

<1 <10 19-27 54

>1 >10  1-25 55

<1 <10 19-28 56

<1 <10 19-29 57

<1 <10 19-30 58

>1 <10 1-1 59

>1 >10 1-3 60

<0.1 <10 12-2  61

<0.1 <10 12-3  62

<0.1 <10 13-2  63

<1 <10 15-5  64

>1 >10 20-2  65

<0.1 <10 12-4  66

<1 <10 19-31 67

<1 <10 19-33 68

<1 <10 19-34 69

<0.1 <10 13-3  70

<1 <10 14-10 71

<0.1 <10 12-8  72

<0.1 <10 12-9  73

<0.1 <10 14-11 74

<1 >10 12-10 75

<0.1 <10 19-35 76

<1 <10 12-5  77

>1 <10 14-12 78

<0.1 <10 13-4  79

<0.1 <10 12-6  80

<0.1 <10 20-3  81

>1 >10 1-4 82

<1 <10 1-5 83

<1 >10 1-6 84

<1 >10 1-7 85

<0.01 <10 1-8 86

<0.01 <10 1-9 87

>1 >10  1-10 88

>1 <10  1-11 89

>1 >10  1-12 90

>1 <10  1-13 91

>1 >10 2-3 92

>1 >10 4-1 93

>1 <10 4-3 94

>1 >10 4-4 95

>1 >10 3-5 96

1 >10 3-1 97

<1 >10 3-3 98

1 <10 3-4 99

>1 <10  1-14 100

<1 >10 3-7 101

<1 >10 19-36 102

<1 <10 19-37 103

>1 >10  1-22 104

>1 >10  1-23 105

<1 <10  1-26 106

>1 >10 1-2 107

>1 >10  1-24 108

>1 >10 3-8 109

<0.1 <10 12-1  110

<1 >10 2-2 111

<1 >10 2-1 112

<1 <10 2-4 113

>1 >10  1-15 114

<1 >10  3-10 115

<1 >10  3-11 116

>1 <10  3-12 117

<1 <10  3-17 118

>1 <10  3-18 119

>1 >10  3-19 120

<1 10 20-4  121

<0.1 <10 20-1  122

>1 <10 5-1 123

<1 nd 4-5 124

<1 <10 12-20 125

<0.1 <10 12-11 126

<0.1 <10 12-12 127

<1 >10 2-5 128

>1 >10 2-6 129

>1 >10 2-7 130

>1 >10 2-8 131

>1 >10 2-9 132

>1 >10  2-10 133

>1 >10  2-11 134

>1 >10  2-12 135

>1 >10  1-16 136

>1 >10  1-17 137

>1 >10  1-18 138

>1 >10  3-13 139

<0.1 <10 12-7  140

<0.1 <10 12-13 141

<0.1 <10 12-14 142

<0.1 <10 12-15 143

>1 <10 13-5  144

<0.1 <10 14-1  145

<1 <10 13-1  146

>1 >10  3-20 147

>1 >10  3-21 148

>1 >10  3-14 149

>1 >10  2-13 150

>1 >10  2-14 151

<1 <10 14-13 152

>1 >10  3-22 153

>1 >10  3-15 154

<1 <10 6-1 155

>1 >10  3-24 156

>1 >10  1-19 157

>1 >10  1-20 158

<1 >10  3-16 159

>1 >10  1-21 160

>1 >10 10-1  161

>1 >10 9-1 162

>1 >10 9-2 163

>1 >10 9-3 164

>1 >10 9-4 165

>1 >10 9-5 166

<1 <10 11-1  167

<1 <10 19-38 168

<1 >10 19-39 169

<0.1 <10 19-40 170

>1 <10 19-41 171

<0.1 <10 19-42 172

<1 >10 3-2 173

<1 >10 3-6 174

>1 <10 3-9 175

>1 <10 4-2 176

>1 >10 6-2 177

<1 >10 7-1 178

>1 >10 8-1 179

>1 >10 9-6 180

<0.1 <10 19-43 181

<1 <10 11b 182

<0.1 <10 11c

Pharmaceutically Acceptable Salts; Stereoisomers; Tautomers

IRE-1α inhibitor compounds include both the free form of the compoundsand the pharmaceutically acceptable salts and stereoisomers thereof.Some of the specific IRE-1α inhibitor compounds described herein are theprotonated salts of amine compounds. The term “free form” refers to theamine compounds in non-salt form. The encompassed pharmaceuticallyacceptable salts not only include the salts described for the specificcompounds disclosed herein, but also all the typical pharmaceuticallyacceptable salts of the free form of IRE-1α inhibitor compounds of theinvention and prodrugs thereof.

The free form of the specific salt compounds described may be isolatedusing techniques known in the art. For example, the free form may beregenerated by treating the salt with a suitable dilute aqueous basesolution such as dilute aqueous NaOH, potassium carbonate, ammonia andsodium bicarbonate. The free forms may differ from their respective saltforms somewhat in certain physical properties, such as solubility inpolar solvents, but the acid and base salts are otherwisepharmaceutically equivalent to their respective free forms for purposesof the invention.

The pharmaceutically acceptable salts of the disclosed IRE-1α inhibitorcompounds can be synthesized from the compounds of this invention whichcontain a basic or acidic moiety by conventional chemical methods.Generally, the salts of the basic compounds are prepared either by ionexchange chromatography or by reacting the free base with stoichiometricamounts or with an excess of the desired salt-forming inorganic ororganic acid in a suitable solvent or various combinations of solvents.Similarly, the salts of the acidic compounds are formed by reactionswith the appropriate inorganic or organic base.

Pharmaceutically acceptable salts of IRE-1α inhibitor compounds includethe conventional non-toxic salts of the compounds as formed by reactinga basic compound with an inorganic or organic acid. For example,conventional non-toxic salts include those derived from inorganic acidssuch as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric,nitric and the like, as well as salts prepared from organic acids suchas acetic, propionic, succinic, glycolic, stearic, lactic, malic,tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic,glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric,toluenesulfonic, benzenesulfonic, methanesulfonic, ethane disulfonic,oxalic, isethionic, trifluoroacetic and the like.

When an IRE-1α inhibitor compound is acidic, suitable pharmaceuticallyacceptable salts include salts prepared form pharmaceutically acceptablenon-toxic bases including inorganic bases and organic bases. Saltsderived from inorganic bases include aluminum, ammonium, calcium,copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous,potassium, sodium, zinc and the like. Particular salts are the ammonium,calcium, magnesium, potassium and sodium salts. Salts derived frompharmaceutically acceptable organic non-toxic bases include salts ofprimary, secondary and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines and basic ionexchange resins, such as arginine, betaine caffeine, choline,N,N1-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol,2-dimethylaminoethanol, ethanolamine, ethylenediamine,N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine,hydrabamine, isopropylamine, lysine, methylglucamine, morpholine,piperazine, piperidine, polyamine resins, procaine, purines,theobromine, triethylamine, trimethylamine tripropylamine, tromethamineand the like. The preparation of the pharmaceutically acceptable saltsdescribed above and other typical pharmaceutically acceptable salts ismore fully described by Berg et al., “Pharmaceutical Salts,” J. Pharm.Sci., 1977:66:1-19.

Some IRE-1α compounds or prodrugs are potentially internal salts orzwitterions, because under physiological conditions a deprotonatedacidic moiety in the compound, such as a carboxyl group, may be anionic,and this electronic charge might then be balanced off internally againstthe cationic charge of a protonated or alkylated basic moiety, such as aquaternary nitrogen atom.

IRE-1α inhibitor compounds or prodrugs thereof may have asymmetriccenters, chiral axes, and chiral planes (as described in: E. L. Elieland S. H. Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons,New York, 1994, pages 1119-1190), and may occur as racemates, racemicmixtures, and as individual diastereomers, with all possible isomers andmixtures thereof, including optical isomers, being included in thepresent invention.

An IRE-1α inhibitor compound or prodrug thereof may be of such a naturethat its constituent atoms are capable of being arranged spatially intwo or more ways, despite having identical bonds. As a consequence, thiscompound exists in the form of stereoisomers. Cis/trans isomerism isonly one type of stereoisomerism. If the stereoisomers are image andmirror image which cannot be superimposed, they are enantiomers whichhave chirality or handedness since one or more asymmetric carbon atomsare present in the structure forming them. Enantiomers are opticallyactive and therefore distinguishable since they rotate the plane ofpolarized light to an equal extent, but in opposite directions.

If two or more asymmetric carbon atoms are present in an IRE-1αcompound, two possible configurations exist at each of these carbonatoms. If two asymmetric carbon atoms are present, four possiblestereoisomers exist, for example. Furthermore, these four possiblestereoisomers can be divided into six possible pairs of stereoisomersthat differ from each other. In order for a pair of molecules with morethan one asymmetric carbon to be enantiomers, they must have differentconfigurations at each asymmetric carbon. Those pairs that do not behaveas enantiomers have a different stereochemical relationship, which isknown as a diastereomeric relationship. Stereoisomers that are notenantiomers are known as diastereoisomers, or, more frequently,diastereomers.

All of these well-known aspects of the stereochemistry of the compoundsof the invention are considered to be part of the present invention. Thepresent invention therefore covers IRE-1α inhibitor compounds which arestereoisomers, and, if these are enantiomers, the individualenantiomers, racemic mixtures of these enantiomers, and artificial, i.e.synthetic, mixtures comprising proportions of these enantiomers whichare different from the proportions of these enantiomers observed in aracemic mixture. If an IRE-1α inhibitor compound has stereoisomers thatare diastereomers, this compound includes the individual diastereomersas well as mixtures of any two or more of these diastereomers in anydesired proportions.

The following is intended to serve for explanation: if a singleasymmetric carbon atom exists in an IRE-1α inhibitor compound thatresults in the (−)(R) and (+)(S) enantiomers thereof, this an IRE-1αinhibitor compound includes all pharmaceutically acceptable salt forms,prodrugs and metabolites thereof which are therapeutically active anduseful for the treatment of or preventing the diseases and conditionsdescribed further herein. If an IRE-1α inhibitor compound exists in theform of (−)(R) and (+)(S) enantiomers, this compound also includes the(+)(S) enantiomer alone or the (−)(R) enantiomer alone if all,substantially all or a predominant share of the therapeutic activityresides in only one of these enantiomers or undesired side effectsreside in only one of these enantiomers. If essentially no differenceexists between the biological properties of the two enantiomers, thiscompound of the invention furthermore includes the (+)(S) enantiomer andthe (−)(R) enantiomer together as a racemic mixture or non-racemicmixture in any desired ratio of corresponding proportions.

The specific biological effects and/or physical and chemical propertiesof a pair or set of enantiomers of an IRE-1α inhibitor compound—ifpresent—may make it obvious to use these enantiomers in certain ratios,for example to form a final therapeutic product. The following isintended to serve for illustration: if a pair of enantiomers exists, theenantiomers can be used in ratios such as 90% (R)-10% (S), 80% (R)-20%(S), 70% (R)-30% (S), 60% (R)-40% (S), 50% (R)-50% (S), 40% (R)-60% (S),30% (R)-70% (S), 20% (R)-80% (S), and 10% (R)-90% (S). After evaluationof the properties of the various enantiomers of an IRE-1α inhibitorcompound—if they exist—the corresponding amount of one or more of theseenantiomers having certain desired properties which form the finaltherapeutic product can be determined in a simple manner.

For IRE-1α inhibitor compounds disclosed herein which may exist astautomers, both tautomeric forms are encompassed within the invention,even though only one tautomeric structure is depicted. For example, acompound such as that below drawn as the keto tautomer includes the enoltautomer, and vice versa, as well as mixtures thereof.

The invention also includes pharmaceutically usable stereoisomers, E/Zisomers, enantiomers, racemates, diastereomers, hydrates, and solvatesof the disclosed compounds. “Solvates” are adductions of inert solventmolecules onto the compounds which form owing to their mutual attractiveforce. Solvates are, for example, monohydrates, dihydrates oralcoholates.

Prodrugs

The invention also provides prodrugs which are metabolized to activeIRE-1α inhibitor compounds after administration. For example, IRE-1αinhibitor compounds disclosed herein can be modified, for example, withalkyl or acyl groups, sugars, or oligopeptides and which are rapidlycleaved in vivo to release the active IRE-1α inhibitor compounds.

Derivatives of the corresponding aromatic alcohols can serve as prodrugsfor aromatic aldehydes because alcohols and aldehydes are metabolicallyinterconvertible, according to the following general scheme:

-   -   Scheline, 1972, Xenobiotica, 2, 227-36.

Examples of prodrugs of aldehydes, ketones, alcohols and otherfunctional groups are described in Wermuth et al., 1996, DesigningProdrugs and Bioprecursors I: Carrier Prodrugs. In The Practice ofMedicinal Chemistry, pp. 672-696; and in Wermuth, 1996, “Preparation ofWater-Soluble Compounds by Covalent Attachment of SolubilizingMoieties,” in Wermuth, ed., The Practice of Medicinal Chemistry, pp.756-776. Other general aldehyde derivatives and alcohol derivatives thatcan perform prodrug functions as well as methods for their preparationare described in Cheronis et al., 1965, Semimicro Qualitative OrganicAnalysis, New York: Interscience, pp. 465-518.

Methods of Preparing IRE-1α Inhibitor Compounds and Prodrugs of theInvention

IRE-1α inhibitor compounds and starting materials for their synthesiscan be prepared by appropriate modification of methods known in the artas described in the literature, for example in standard works such asHouben-Weyl, Methoden der organischen Chemie, Georg-Thieme-Verlag,Stuttgart. Methods may also be found by computer search in The MDL®CrossFire Beilstein database, in which the reaction domain details thepreparation of substances. See also the specific Examples, below.

Pharmaceutical Preparations

Any of the IRE-1α inhibitor compounds and prodrugs disclosed herein canbe formulated as pharmaceuticals using methods well known in the art.Pharmaceutical formulations of the invention typically comprise at leastone IRE-1α inhibitor compound or prodrug thereof mixed with a carrier,diluted with a diluent, and/or enclosed or encapsulated by an ingestiblecarrier in the form of a capsule, sachet, cachet, paper, or othercontainer, or by a disposable container such as an ampoule.

A carrier or diluent can be a solid, semi-solid, or liquid material.Some examples of diluents or carriers which can be employed in thepharmaceutical compositions of the present invention are lactose,dextrose, sucrose, sorbitol, mannitol, propylene glycol, liquidparaffin, white soft paraffin, kaolin, microcrystalline cellulose,calcium silicate, silica polyvinylpyrrolidone, cetostearyl alcohol,starch, gum acacia, calcium phosphate, cocoa butter, oil of theobroma,arachis oil, alginates, tragacanth, gelatin, methyl cellulose,polyoxyethylene sorbitan monolaurate, ethyl lactate,propylhydroxybenzoate, sorbitan trioleate, sorbitan sesquioleate, andoleyl alcohol.

Pharmaceutical compositions of the invention can be manufactured bymethods well known in the art, including conventional mixing,dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping, or lyophilizing processes.

For injection, the IRE-1α inhibitor compounds of the invention can beformulated in aqueous solutions, preferably in physiologicallycompatible buffers such as Hanks's solution, Ringer's solution, orphysiological saline buffer. For transmucosal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art. If desired, any of theIRE-1α inhibitor compounds or prodrugs thereof disclosed herein can beprovided in a pyrogen-free pharmaceutically acceptable vehicle.

For oral administration, an IRE-1α inhibitor compound or prodrug thereofcan be combined with pharmaceutically acceptable carriers or vehicleswhich enable the IRE-1α inhibitor compound or prodrug thereof to beformulated as tablets, pills, dragees, capsules, liquids, gels, syrups,slurries, suspensions and the like. Fillers can be used, such asgelatin, sugars (e.g., lactose, sucrose, mannitol, or sorbitol),cellulose preparations (e.g., maize starch, wheat starch, rice starch,potato starch, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose), and/orpolyvinylpyrrolidone (PVP). If desired, disintegrating agents can beadded, such as the cross-linked polyvinyl pyrrolidone, agar, or alginicacid or a salt thereof, such as sodium alginate.

Dragee cores can be provided with suitable coatings. For this purpose,concentrated sugar solutions can be used, which can optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments can be added to thetablets or dragee coatings for identification.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with a fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, an IRE-1α inhibitor compound or prodrug thereof can bedissolved or suspended in a suitable liquid, such as fatty oils, liquidparaffin, or liquid polyethylene glycols. In addition, stabilizers canbe added. All formulations for oral administration preferably are indosages suitable for such administration.

For buccal administration, the compositions can take the form of tabletsor lozenges formulated in conventional manners.

For administration by inhalation, pharmaceutical preparations of theinvention can be delivered in the form of an aerosol sprays from apressurized pack or a nebulizer, with the use of a suitable propellant,e.g., dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. Ifdesired, a valve can be used to deliver a metered amount. Capsules andcartridges of, e.g., gelatin for use in an inhaler or insufflator, canbe formulated containing a powder mix of an IRE-1α inhibitor compound orprodrug thereof and a suitable powder base, such as lactose or starch.

IRE-1α inhibitor compounds or prodrugs thereof can be formulated forparenteral administration by injection, e.g., by bolus injection orcontinuous infusion. Formulations for injection can be presented in unitdosage form, e.g., in ampoules or in multi-dose containers, with anadded preservative. The compositions can take such forms as suspensions,solutions, or emulsions in oily or aqueous vehicles, and can containformulatory agents such as suspending, stabilizing, and/or dispersingagents.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of an IRE-1α inhibitor compound or prodrug thereof.Additionally, a suspension of an IRE-1α inhibitor compound or prodrugthereof can be prepared as an appropriate oily injection suspension.Suitable lipophilic solvents or vehicles include fatty oils such assesame oil, synthetic fatty acid esters, such as ethyl oleate ortriglycerides, or liposomes. Aqueous injection suspensions can containsubstances which increase the viscosity of the suspension, such assodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, thesuspension can also contain suitable stabilizers or agents whichincrease the solubility of an IRE-1α inhibitor compound or prodrugthereof to allow for the preparation of highly concentrated solutions.

Alternatively, an IRE-1α inhibitor compound or prodrug thereof can be inpowder form for constitution with a suitable vehicle, e.g., sterilepyrogen-free water, before use.

IRE-1α inhibitor compounds or prodrugs thereof can also be formulated inrectal compositions such as suppositories or retention enemas, e.g.,containing conventional suppository bases such as cocoa butter or otherglycerides.

In addition to the formulations described previously, an IRE-1αinhibitor compound or prodrug thereof can also be formulated as a depotpreparation. Such long-acting formulations can be administered byimplantation (for example, subcutaneously or intramuscularly) or byintramuscular injection. Thus, for example, an IRE-1α inhibitor compoundor prodrug thereof can be formulated with suitable polymeric orhydrophobic materials (for example as an emulsion in an acceptable oil)or ion exchange resins, or as sparingly soluble derivatives, forexample, as a sparingly soluble salt.

The pharmaceutical compositions also can comprise suitable solid or gelphase carriers or excipients. Examples of such carriers or excipientsinclude but are not limited to, calcium carbonate, calcium phosphate,various sugars, starches, cellulose derivatives, gelatin, and polymerssuch as polyethylene glycols.

In addition to the common dosage forms set out above, an IRE-1αinhibitor compound or prodrug thereof can be administered by acontrolled release means and/or delivery device, including ALZET®osmotic pumps (Alza Corporation). Suitable delivery devices aredescribed in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123;3,944,064; and 4,008,719.

Therapeutic Methods

IRE-1α inhibitor compounds or prodrugs thereof can be administered to apatient, preferably a human patient, in pharmaceutical preparations asdisclosed above, preferably with a pyrogen-free pharmaceuticallyacceptable vehicle, at doses effective to treat or ameliorate a symptomof a disorder associated with the unfolded protein response.

Disorders Associated with UPR

A fine balance exists between a cell's life and death depending on howprotein folding stress is managed by the cell (proteostasis). Imbalancesin proteostasis lead to many metabolic, oncological, neurodegenerative,inflammatory, cardiovascular disorders and infectious disease (Balch etal., Science 319, 916, 2008). The UPR relates specifically to theproteostasis of the endoplasmic reticulum where all secreted andmembrane proteins are translated, folded and processed for delivery totheir individual site of action. Therefore, activation of the UPRenhances protein folding in the ER allowing the cell to survive. Ifprotein folding stress is not managed in the ER, the cells will initiateapoptosis.

Protein folding stress may be a natural hallmark of the type of cell forexample insulin secreting β-islet cells or antibody secreting plasmacells. In both cases, the cell has fine tuned the machinery to deal withthe stress by activating the UPR. Depending on the disease type, it maybe therapeutically beneficial to induce or inhibit the UPR. For example,in type II diabetes or Alzheimer's disease, it may be therapeuticallybeneficial to activate the UPR in such a way where β-islet cells survivethe stress of over producing insulin or neurons survive the apoptoticeffects due to unfolded aggregates of β-amyloid protein. Diseases suchas cancer, inflammation, and viral infection may be therapeuticallymodulated by inhibition of the UPR. In these types of conditions,cellular survival due to corruption of the UPR may be impacted. Proteinfolding in the ER is negatively impacted by such conditions in the tumormicroenvironment as hypoxia, glucose starvation, amino acid deprivation,acidosis and mutant malfolded and oncgenic proteins. Additionallychemo-, bio-, and radiotherapy can lead to protein folding stress. Itmay be possible to induce apoptosis in these conditions by inhibitingthe anti-apoptotic effects of the UPR. Myeloma derived from neoplasticantibody secreting plasma cells provides an example of a condition inwhich this approach can be applied.

Lastly, enveloped viruses must use and corrupt this system to ensureproduction of progeny from infected cells. Viruses often produce vastquantities of viral membrane glycoproteins which are folded and modifiedin the ER. Therefore, activation of the UPR by the virus for thispurpose as a survival mechanism is entirely conceivable. It is thereforelogical that inhibition of the UPR during viral infection can impact theoutcome of the disease in a beneficial way.

Only specialized secretory cells and diseased cells activate the UPR fortheir own benefit. Most cells are not under such protein folding stressand therefore would not be impacted by a UPR inhibitor. Thus, “disordersassociated with the UPR” as used herein means conditions for whichpathogenesis can be advantageously impacted by inhibition of the UPR. Invarious embodiments of the invention such inhibition of the UPR isaccomplished through inhibition of IRE-1α.

In some embodiments the IRE-1α inhibitor compounds or prodrugs thereofare useful to treat or ameliorate a symptom of a B cell autoimmunedisease, certain cancers, and infections of enveloped viruses that usethe endoplasmic reticulum as a viral factory for expressing viralsurface and spike proteins for budding and infection. IRE-1α inhibitorsand prodrugs thereof can be used as single agents or in combinationtherapies, as described below.

B cell autoimmune diseases which can be treated include, but are notlimited to, Addison's disease, antiphospholipid syndrome, aplasticanemia, autoimmune hemolytic anemias, autoimmune hepatitis, autoimmunehypophysitis, autoimmune lymphoproliferative disorders, autoimmunemyocarditis, Churg-Strauss syndrome, epidermolysis bullosa acquisita,giant cell arteritis, Goodpasture's syndrome, Graves' disease,Guillain-Barré syndrome, Hashimoto's thyroiditis, idiopathicthrombocytopenic purpura, IgA nephropathy, myasthenia gravis, pemphigusfoliaceous, pemphigus vulgaris, polyarteritis nodosa,polymyositis/dermatomyositis, rheumatoid arthritis, scleroderma,Sjögren's syndrome, systemic lupus erythematosus, Takayasu's arteritis,and Wegener's granulomatosis.

Cancers which can be treated include solid tumors, such as tumors of thebreast, bone, prostate, lung, adrenal gland (e.g., adrenocorticaltumors), bile duct, bladder, bronchus, nervous tissue (includingneuronal and glial tumors), gall bladder, stomach, salivary gland,esophagus, small intestine, cervix, colon, rectum, liver, ovary,pancreas, pituitary adenomas, and secretory adenomas. Methods of theinvention are particularly useful for treating drug- orradiation-resistant solid tumors.

Cancers of the blood (e.g., lymphomas and leukemias) also can be treatedincluding, but not limited to, multiple myeloma, Hodgkin's lymphoma,non-Hodgkin's lymphomas (e.g., cutaneous T cell lymphomas such as Sezarysyndrome and Mycosis fungoides, diffuse large cell lymphoma, HTLV-1associated T cell lymphoma, nodal peripheral T cell lymphoma, extranodalperipheral T cell lymphoma, central nervous system lymphoma, andAIDS-related lymphoma). Leukemias include acute and chronic types ofboth lymphocytic and myelogenous leukemia (e.g, acute lymphocytic orlymphoblastic leukemia, acute myelogenous leukemia, acute myeloidleukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, Tcell prolymphocytic leukemia, adult T cell leukemia, and hairy cellleukemia). Monoclonal gammopathy of undetermined significance (MGUS),the precursor of myeloma, also can be treated.

Viral infections which can be treated include infections of envelopedviruses which use the unfolded protein response pathway when theyreplicate and form infectious progeny (e.g., measles, pox viruses,Ebola, etc.). Infections also include those of Epstein Barr virus (EBV),cytomegalovirus (CMV), Flaviviruses (e.g., Japanese Encephalitis Virusand West Nile Virus), and Hepatitis C virus (HCV).

Combination Therapies

Various types of physiological stress induce the unfolded proteinresponse including, but not limited to, hypoxia, nutrient starvation,acidosis, and genetic damage resulting in mutant or over-expressedmisfolded proteins (oncogenic stress). One or more of these conditionsare manifest in cancer cells, which may in part be mediated by themicroenviroment of the tumor. It is likely the cytoprotective arm of theunfolded protein response (UPR) plays an anti-apototic role in tumorsurvival. In addition, bio- and chemotherapeutic drugs and radiationtreatments may further impact the protein folding and degradation cyclein the ER thereby inducing the UPR as a protective resistance mechanism.Patients succumb to cancer because either the tumor is resistant toconventional therapies or returns in a resistant form after an initialresponse to treatment and, therefore, new treatments and treatmentcombinations are needed.

Angiogenesis inhibitors block tumor growth by inhibiting new bloodvessel formation, a process that would enhance the stress effects of thetumor microenvironment. A promising approach to further reduce tumorburden would be to administer anti-angiogenesis agents in combinationwith IRE-1α/XBP-1 inhibitors to obtain a similar effect as thatdemonstrated by RNAi knockdown of GRP78, the major chaperone of the ERand target of XBP-1s (Dong et al., Cancer Res. 2007 Jul. 15;67(2):6700-7). In addition, IRE-1α itself regulates angiogensis byinfluencing the expression of VEGF.

Proteasome inhibitors and Hsp90 inhibitors are thought to act in part byblocking protein degradation and folding, respectively, inducingapoptosis (Davenport et al., Blood 2007 Oct. 1; 110(7):2641-9). Althoughit is clear that Hsp90 inhibitors induce XBP-1 splicing and activationof the UPR, it is less clear that proteasome inhibitors activate IRE-1α.Current scientific literature suggest that IRE-1α is not or is onlyminimally activated by proteasome inhibitors, such as bortezomib orMG-132 (Davenport et al., Blood 2007 Oct. 1; 110(7):2641-9).

Interference with UPR may sensitize cancer cells to variouschemotherapeutics that elevate the cellular stress. Combinationtherapies which include IRE-1α inhibitors may become important therapiesin conjunction with current and future standard of care in cancer.

Although the level of activation IRE-1α in solid tumors is currently notknown, clearly, induction of the UPR in patient biopsies of drugresistant tumors is evidenced by induction of GRP78 (Moenner et al.,Cancer Res. 2007 Nov. 15; 67(22):10631-4; Lee, Cancer Res. 2007 Apr. 15;67(6e):3496-9).

Inhibition of XBP-1 splicing may have a greater effect than anticipatedas the un-spliced form of XBP-1 may act as a dominant negative to XBP-1and ATF-6 transcriptional activity. Further inhibitors which block theRNAse activity but not kinase activity of IRE-1α may have the addedbenefit of signaling through the JNK pathway, a signal that can havepro-apoptotic consequences.

In some embodiments an IRE-1α inhibitor compound or prodrug thereof isadministered in combination with a therapeutic agent that induces orup-regulates IRE-1α expression (e.g., Hsp90 and or HDAC inhibitors, bothof which induce IRE-1α activation and XBP-1 splicing) or a therapeuticagent which is less effective when IRE-1α is expressed (e.g., 17-AAG(TANESPIMYCIN® and suberoylanilide hydroxamic acid (SAHA)).

In some embodiments an IRE-1α inhibitor compound or prodrug thereof isadministered in combination with a cancer therapeutic agent, for exampleradiation therapy or a cancer therapeutic agent (e.g., achemotherapeutic agent or a biotherapeutic agent) as described below.The cancer therapeutic agent can be administered separately or togetherwith the IRE-1α inhibitor compound. The cancer therapeutic agent can beadministered at essentially the same time as the IRE-1α inhibitorcompound or can be administered either before or after the IRE-1αinhibitor compound.

Cancer therapeutic agents which can be used according to the inventioninclude, but are not limited to, agents in the following categories(which may overlap):

-   -   a. proteasome inhibitors, such as bortezomib        ([(1R)-3-methyl-1-[[(2S)-1-oxo-3-phenyl-2-[(pyrazinylcarbonyl)amino]propyl]amino]butyl]        boronic acid; MG-341; VELCADE®), MG-132        (N-[(phenylmethoxy)carbonyl]-L-leucyl-N-[(1S)-1-formyl-3-methylbutyl]-L-leucinamide);    -   b. antimetabolites, such as:        -   i. pyrimidine analogs (e.g., 5-fluorouracil, floxuridine,            capecitabine, gemcitabine and cytarabine);        -   ii. purine analogs,        -   iii. folate antagonists and related inhibitors (e.g.,            mercaptopurine, thioguanine, pentostatin and            2-chlorodeoxyadenosine [cladribine]);        -   iv. folic acid analogs (e.g., methotrexate);    -   c. antimitotic agents, including:        -   i. natural products such as vinca alkaloids (e.g.,            vinblastine, vincristine, and vinorelbine);        -   ii. alkylating agents such as nitrogen mustards (e.g.,            mechlorethamine, cyclophosphamide and analogs, melphalan,            chlorambucil), ethylenimines and methylmelamines (e.g.,            hexamethylmelamine and thiotepa), alkyl sulfonates-busulfan,            nitrosoureas (e.g., carmustine (BCNU) and analogs,            streptozocin), trazenes—dacarbazinine (DTIC);    -   d. microtubule disruptors such as taxane (paclitaxel,        docetaxel), vincristin, vinblastin, nocodazole, epothilones and        navelbine, and epidipodophyllotoxins (e.g., teniposide);    -   e. DNA damaging agents, such as actinomycin, amsacrine,        anthracyclines, bleomycin, busulfan, camptothecin, carboplatin,        chlorambucil, cisplatin, cyclophosphamide, Cytoxan,        dactinomycin, daunorubicin, docetaxel, doxorubicin, epirubicin,        hexamethylmelamineoxaliplatin, iphosphamide, melphalan,        merchlorethamine, mitomycin, mitoxantrone, nitrosourea,        paclitaxel, plicamycin, procarbazine, teniposide,        triethylenethiophosphoramide and etoposide (VP 16);    -   f. antibiotics, such as dactinomycin (actinomycin D),        daunorubicin, doxorubicin (adriamycin), idarubicin,        anthracyclines, mitoxantrone, bleomycins, plicamycin        (mithramycin) and mitomycin;    -   g. enzymes, such as L-asparaginase;    -   h. antiplatelet agents;    -   i. platinum coordination complexes (e.g., cisplatin,        carboplatin), procarbazine, hydroxyurea, mitotane,        aminoglutethimide;    -   j. hormones, hormone analogs (e.g., estrogen, tamoxifen,        goserelin, bicalutamide, nilutamide);    -   k. aromatase inhibitors (e.g., letrozole, anastrozole);    -   l. anticoagulants (e.g., heparin, synthetic heparin salts and        other inhibitors of thrombin);    -   m. fibrinolytic agents (such as tissue plasminogen activator,        streptokinase and urokinase), aspirin, COX-2 inhibitors,        dipyridamole, ticlopidine, clopidogrel, abciximab;    -   n. antimigratory agents;    -   o. antisecretory agents (e.g., breveldin); immunosuppressives        (e.g., cyclosporine, tacrolimus (FK-506), sirolimus (rapamycin),        azathioprine, mycophenolate mofetil);    -   p. anti-angiogenic compounds (e.g., TNP-470, genistein) and        growth factor inhibitors (e.g., vascular endothelial growth        factor (VEGF) inhibitors, fibroblast growth factor (FGF)        inhibitors, epidermal growth factor (EGF) inhibitors);    -   q. angiotensin receptor blockers;    -   r. nitric oxide donors;    -   s. anti-sense oligonucleotides;    -   t. antibodies (e.g., trastuzumab (HERCEPTIN®), AVASTIN®,        ERBITUX®);    -   u. cell cycle inhibitors and differentiation inducers (e.g.,        tretinoin);    -   v. mTOR (mammalian target of rapamycin) inhibitors (e.g.,        everolimus, sirolimus);    -   w. topoisomerase inhibitors (e.g., doxorubicin (adriamycin),        amsacrine, camptothecin, daunorubicin, dactinomycin, eniposide,        epirubicin, etoposide, idarubicin, irinotecan (CPT-11) and        mitoxantrone, topotecan, irinotecan);    -   x. corticosteroids (e.g., cortisone, dexamethasone,        hydrocortisone, methylprednisolone, prednisone, and        prenisolone);    -   y. growth factor signal transduction kinase inhibitors;    -   z. mitochondrial dysfunction inducers;    -   aa. caspase activators; and    -   bb. chromatin disruptors.

In some embodiments the cancer therapeutic agent is selected from thegroup consisting of alemtuzumab, aminoglutethimide, amsacrine,anastrozole, asparaginase, beg, bevacizumab, bicalutamide, bleomycin,bortezomib, buserelin, busulfan, campothecin, capecitabine, carboplatin,carmustine, CeaVac, cetuximab, chlorambucil, cisplatin, cladribine,clodronate, colchicine, cyclophosphamide, cyproterone, cytarabine,dacarbazine, daclizumab, dactinomycin, daunorubicin, dienestrol,diethylstilbestrol, docetaxel, doxorubicin, edrecolomab, epirubicin,epratuzumab, erlotinib, estradiol, estramustine, etoposide, exemestane,filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone,flutamide, gemcitabine, gemtuzumab, genistein, goserelin, huJ591,hydroxyurea, ibritumomab, idarubicin, ifosfamide, IGN-101, imatinib,interferon, irinotecan, ironotecan, letrozole, leucovorin, leuprolide,levamisole, lintuzumab, lomustine, MDX-210, mechlorethamine,medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna,methotrexate, mitomycin, mitotane, mitoxantrone, mitumomab, nilutamide,nocodazole, octreotide, oxaliplatin, paclitaxel, pamidronate,pentostatin, pertuzumab, plicamycin, porfimer, procarbazine,raltitrexed, rituximab, streptozocin, sunitinib, suramin, tamoxifen,temozolomide, teniposide, testosterone, thalidomide, thioguanine,thiotepa, titanocene dichloride, topotecan, tositumomab, trastuzumab,tretinoin, vatalanib, vinblastine, vincristine, vindesine, andvinorelbine.

Routes of Administration

Pharmaceutical preparations of the invention can be administered locallyor systemically. Suitable routes of administration include oral,pulmonary, rectal, transmucosal, intestinal, parenteral (includingintramuscular, subcutaneous, intramedullary routes), intranodal,intrathecal, direct intraventricular, intravenous, intraperitoneal,intranasal, intraocular, transdermal, topical, and vaginal routes. Asdescribed in more detail above, dosage forms include, but are notlimited to, tablets, troches, dispersions, suspensions, suppositories,solutions, capsules, creams, patches, minipumps and the like. Targeteddelivery systems also can be used (for example, a liposome coated withtarget-specific antibody).

Dosage

A pharmaceutical composition of the invention comprises at least oneactive ingredient (an IRE-1α inhibitor compound or prodrug thereof) in atherapeutically effective dose. A “therapeutically effective dose” isthe amount of an IRE-1α inhibitor compound or prodrug thereof which,when administered to a patient over a treatment period, results in ameasurable improvement in a characteristic of the disease being treated(e.g., improved laboratory values, retarded development of a symptom,reduced severity of a symptom, or improved levels of an appropriatebiological marker).

Determination of therapeutically effective doses is well within thecapability of those skilled in the art. A therapeutically effective doseinitially can be estimated from in vitro enzyme assays, cell cultureassays, and/or animal models. For example, a dose can be formulated inan animal model to achieve a circulating concentration range at least asconcentrated as the IC₅₀ as determined in an in vitro enzyme assay or ina cell culture (i.e., the concentration of the test compound whichachieves a half-maximal inhibition of IRE-1α activity). Such informationcan be used to more accurately determine useful doses in humans. See theFDA guidance document “Guidance for Industry and Reviewers Estimatingthe Safe Starting Dose in Clinical Trials for Therapeutics in AdultHealthy Volunteers” (HFA-305), which provides an equation for use incalculating a human equivalent dose (HED) based on in vivo animalstudies.

Appropriate animal models for the relevant diseases are known in theart. See, e.g., Lupus. 1996 Oct.; 5(5b):451-5 (antiphospholipidsyndrome); Blood. 1974 July; 44(1):49-56 (aplastic anemia);Autoimmunity. 2001; 33(5):265-74 (autoimmune hypophysitis); Methods.2007 January; 41(1):118-22 (autoimmune myocarditis); Clin Exp Rheumatol.2003 Nov.-Dec.; 21(6 Suppl 32):S55-63 (Churg-Strauss syndrome, Wegener'sgranulomatosis); J Clin Invest. 2005 April; 115(5):870-8 (epidermolysisbullosa acquisita); Circulation. 2005 Jun. 14; 111(23):3135-40. Epub2005 Jun. 6 (giant cell arteritis; Takayusu's arteritis); Int JImmunopathol Pharmacol. 2005 Oct.-Dec.; 18(5):701-8 (IgA nephropathy);Vet Rec. 1984 May 12; 114(19):479 (pemphigus foliaceous); J.Neuroimmunol. 98, 130-35, 1999 (polymyositis); Am. J. Pathol. 120,323-25, 1985 (dermatomyositis); Cell. Mol. Immunol. 2, 461-65, 2005(myasthenia gravis); Arthritis Rheum. 50, 3250-59, 2004 (lupuserythymatosus); Clin. Exp. Immunol. 99, 294-302, 1995 (Grave's disease);J. Clin. Invest. 116, 961-973, 2006 (rheumatoid arthritis); Exp MolPathol. 77, 161-67, 2004 (Hashimoto's thyroiditis); Rheumatol. 32,1071-75, 2005 (Sjögren's syndrome); Brain Pathol. 12, 420-29, 2002(Guillain-Barré syndrome); Vet. Pathol. 32, 337-45, 1995 (polyarteritisnodosa); Immunol. Invest. 3, 47-61, 2006 (pemphigus vulgaris); Arch.Dermatol. Res. 297, 333-44, 2006 (scleroderma); J. Exp. Med. 191,899-906, 2000 (Goodpasture's syndrome); Clin. Exp. Immunol. 99, 294-302,1995 (Grave's disease); J. Clin. Invest. 91, 1507-15, 1993 (membranousnephropathy); J. Immunol. 169, 4889-96, 2002 (autoimmune hepatitis);Surgery 128, 999-1006, 2000 (Addison's disease); Eur. J. Immunol. 32,1147-56, 2002 (autoimmune hemolytic anemia); and Haematologica 88,679-87, 2003 (autoimmune thrombocytopenic purpura).

LD₅₀ (the dose lethal to 50% of the population) and the ED₅₀ (the dosetherapeutically effective in 50% of the population) can be determined bystandard pharmaceutical procedures in cell cultures and/or experimentalanimals. Data obtained from cell culture assays or animal studies can beused to determine initial human doses. As is known in the art, thedosage may vary depending upon the dosage form and route ofadministration used.

Usual dosages for systemic administration to a human patient range from1 μg/kg to 100 mg/kg (e.g., 1-10 μg/kg, 20-80 μg/kg, 5-50 μg/kg, 75-150μg/kg, 100-500 μg/kg, 250-750 μg/kg, 500-1000 μg/kg, 1-10 mg/kg, 5-50mg/kg, 25-75 mg/kg, 50-100 mg/kg, 5 mg/kg, 20 mg/kg, or 50 mg/kg). Insome embodiments, the treatment schedule can require that a plasmaconcentration of an IRE-1α inhibitor compound be maintained for a periodof time (e.g., several days or a week) and then allowed to decay byceasing administration for a period of time (e.g., 1, 2, 3, or 4 weeks).The amount of composition administered will, of course, be dependent onthe subject being treated, on the subject's weight, the severity of thedisorder, the manner of administration and the judgment of theprescribing physician.

All patents, patent applications, and references cited in thisdisclosure are expressly incorporated herein by reference. The abovedisclosure generally describes the present invention. A more completeunderstanding can be obtained by reference to the following specificexamples, which are provided for purposes of illustration only and arenot intended to limit the scope of the invention.

EXAMPLES

The analytical LC/MS method used in Examples 1-20 employed an Agilent1200 with Variable Wavelength detector extracted at 220 nm and Agilent6140 Single quadrupole mass spectrometer. The HPLC column was a ZorbaxSB-C18, 3.5 μm, 2.1 mm×30 mm, maintained at 40° C. The HPLC Gradient was0.4 mL/min, 95:5:0.1 water:acetonitrile:formic acid for 0.1 min then to5:95:0.1 water:acetonitrile:formic acid in 3.9 min, maintaining for 0.5min.

Example 1 Synthesis of2-Hydroxy-6-(4-methyl-piperazin-1-yl)-naphthalene-1-carbaldehydehydrochloride 1-1

2-Hydroxy-6-(4-methyl-piperazin-1-yl)-naphthalene-1-carbaldehyde 1-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde (1a, WO2008154484) (100 mg,0.4 mmol), 1-methyl-piperazine (44 mg, 0.44 mmol), sodium tert-butoxide(84 mg, 0.88 mmol), tris-(dibenzylideneacetone)dipalladium(0) (25 mg, 28mol), (2-biphenyl)di-tert-butylphosphine (18 mg, 26 μmol) were dissolvedin 12 mL of dry dioxane. The resulted tan slurry was heated to 100° C.for 1 h. The reaction mixture was evaporated and partitioned between 20mL of chloroform and 20 mL of water. The pH of the aqueous phase wasadjusted to neutral with acetic acid then was separated, and extractedwith another 20 mL portion of chloroform. The combined organic phaseswere dried over sodium sulfate, filtered and evaporated. The resultedsolid material was purified by chromatography with chloroform as eluent.The obtained crude product was triturated with diethyl ether to afford1-1 (50 mg, 19 mmol, 46%).

LC/MS ESI: M+H=271, Rt: 2.70 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.72(br. s., 1H), 11.03 (br. s., 1H), 10.76 (s, 1H), 8.85 (d, J=9.3 Hz, 1H),7.99 (d, J=8.8 Hz, 1H), 7.49 (dd, J=9.4, 2.6 Hz, 1H), 7.32 (d, J=2.8 Hz,1H), 7.23 (d, J=9.0 Hz, 1H), 3.84-3.97 (m, 2H), 3.45-3.58 (m, 2H), 3.17(d, J=9.3 Hz, 4H), 2.81 (d, J=4.8 Hz, 3H).

The following compounds were made by the above procedure:

No. MW M + H Rt 1-1 

270.3 271 2.70 1-2 

284.4 285 2.64 1-3 

241.3 242 4.05 1-4 

333.4 334 2.97 1-5 

340.4 341 2.69 1-6 

298.3 299 3.23 1-7 

312.4 313 2.75 1-8 

298.4 299 2.64 1-9 

353.5 354 2.43 1-10

270.3 271 2.63 1-11

257.3 258 3.60 1-12

353.5 354 2.55 1-13

340.4 341 2.72 1-14

298.4 299 2.65 1-15

339.4 340 2.01 1-16

284.4 285 2.64 1-17

326.4 327 2.65 1-18

324.4 325 2.83 1-19

298.3 299 3.27 1-20

312.4 313 3.67 1-21

298.4 299 2.67 1-22

333.4 334 2.92 1-23

241.3 242 4.24 1-24

326.4 327 2.66 1-25

257.3 258 3.20 1-26

283.1 284

Example 2 Synthesis of2-Hydroxy-6-[3-(morpholine-4-carbonyl)-pyrrolidin-1-yl]-naphthalene-1-carbaldehyde2-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (150 mg, 0.6 mmol),morpholin-4-yl-pyrrolidin-3-yl-methanone (158 mg, 0.72 mmol),sodium-tert-butoxide (207 mg, 2.16 mmol),tris-(dibenzylideneacetone)dipalladium(0) (38 mg, 41 mol),(2-biphenyl)di-tert-butylphosphine (25 mg, 84 μmol) were dissolved in 18mL of dry dioxane. The resulted tan slurry was heated to 100° C. for 3h. The reaction mixture was evaporated and partitioned between 30 mL ofchloroform and 30 mL of water. The pH of the aqueous phase was adjustedto neutral with acetic acid then was separated, and extracted withanother 30 mL portion of chloroform. The combined organic phases weredried over sodium sulfate, filtered and evaporated. The resulted solidmaterial was purified by chromatography with chloroform as eluent. Theobtained crude product was triturated with diethyl ether to afford 2-1(108 mg, 31 mmol, 51%).

LC/MS ESI: M+H=355, Rt: 3.43 min; ¹H NMR (400 MHz, CDCl₃) δ ppm 12.80(s, 1H), 10.75 (s, 1H), 8.21 (d, J=9.3 Hz, 1H), 7.81 (d, J=9.0 Hz, 1H),7.08 (dd, J=9.3, 2.5 Hz, 1H), 7.05 (d, J=9.0 Hz, 1H), 6.78 (d, J=2.5 Hz,1H), 3.58-3.78 (m, 10H), 3.52-3.58 (m, 1H), 3.45-3.51 (m, 1H), 3.35-3.45(m, 1H), 2.34-2.46 (m, 1H), 2.21-2.33 (m, 1H).

The following compounds were made by the above procedure:

No. MW M + H Rt 2-1 

354.4 355 3.43 2-2 

368.4 369 2.91 2-3 

381.5 382 2.41 2-4 

367.4 368 2.81 2-5 

298.3 299 3.21 2-6 

312.4 313 3.45 2-7 

352.4 353 3.92 2-8 

388.5 389 4.09 2-9 

298.3 299 2.28 2-10

312.4 313 3.55 2-11

352.4 353 3.96 2-12

388.5 389 4.14 2-13

354.4 355 3.48 2-14

367.4 368 2.74

Example 3 Synthesis of2-Hydroxy-6-(4-pyridin-2-ylmethyl-piperazin-1-yl)-naphthalene-1-carbaldehyde3-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (83 mg, 0.32 mmol),1-pyridin-2-ylmethyl-piperazine hydrochloride (84 mg, 0.39 mmol),sodium-tert-butoxide (138 mg, 1.44 mmol),tris-(dibenzylideneacetone)dipalladium(0) (20 mg, 22 μmol),(2-biphenyl)di-tert-butylphosphine (25 mg, 47 μmol) were dissolved in 8mL of dry dioxane. The resulted tan slurry was heated to 100° C. for 3h. The reaction mixture was evaporated and partitioned between 15 mL ofchloroform and 15 mL of water. The pH of the aqueous phase was adjustedto neutral with acetic acid then was separated, and extracted withanother 15 mL portion of chloroform. The combined organic phases weredried over sodium sulfate, filtered and evaporated. The resulted solidmaterial was purified by chromatography with chloroform/methanol (98/2)as eluent. The obtained crude product was suspended in 5 mL of HCl indioxane, filtered and washed with diethyl ether to afford 3-1 (26 mg,0.7 mmol, 21%).

LC/MS ESI: M+H=348, Rt: 2.75 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.64(br. s., 1H), 10.76 (s, 1H), 8.85 (d, J=9.3 Hz, 1H), 8.71 (d, J=4.3 Hz,1H), 7.94-8.02 (m, 2H), 7.74 (d, J=7.8 Hz, 1H), 7.53 (dd, J=7.2, 4.9 Hz,1H), 7.47 (dd, J=9.3, 2.5 Hz, 1H), 7.30 (d, J=2.5 Hz, 1H), 7.24 (d,J=9.0 Hz, 1H), 4.58 (s, 2H), 3.58 (br. s., 4H), 3.44 (d, J=4.0 Hz, 4H).

The following compounds were made by the above procedure:

No. MW M + H Rt 3-1 

347.4 348 2.76 3-2 

374.5 375 3.01 3-3 

360.5 361 3.07 3-4 

360.5 361 3.16 3-5 

346.4 345 2.99 3-6 

375.5 376 2.70 3-7 

347.4 348 2.65 3-8 

361.4 362 2.99 3-9 

375.5 376 2.45 3-10

361.4 362 2.69 3-11

361.4 362 2.85 3-12

361.4 362 2.60 3-13

339.4 338 (M − H) 2.74 3-14

389.5 390 2.97 3-15

346.4 347 3.06 3-16

389.5 390 2.95 3-17

324.4 325 2.86 3-18

339.4 340 2.76 3-19

375.5 376 2.78 3-20

360.5 361 3.04 3-21

361.4 362 2.69 3-22

360.5 361 3.16 3-23

346.4 347 3.02 3-24

361.4 362 2.78

Example 4 Synthesis of2-Hydroxy-6-(4-methanesulfonyl-piperazin-1-yl)-naphthalene-1-carbaldehyde4-1

4-Methanesulfonyl-piperazine-1-carboxylic acid tert-butyl ester 4a

Piperazine-1-carboxylic acid tert-butyl ester (562 mg, 3.02 mmol) andtriethylamine (915 mg, 9.06 mmol) were dissolved in 30 mLdichloroethane, cooled to 0° C., and methanesulfonyl chloride (257 μL,3.32 mmol) was added dropwise, and the mixture was stirred in thecooling bath for 2 h. Then the mixture was extracted with 5% citric acidand brine. The organic layers was dried over sodium sulfate, filteredand evaporated to afford as a white solid (644 mg, 2.44 mmol, 80%).

1-Methanesulfonyl-piperazine hydrochloride 4b

4-Methanesulfonyl-piperazine-1-carboxylic acid tert-butyl ester (640 mg,2.42 mmol was dissolved in ethyl acetate (20 mL) and ethyl acetatecontaining HCl was added to the solution at 0° C. and let to reach roomtemperature. After 3 h stirring, the suspension was filtered and washedwith diethyl ether to obtain 4b (420 mg, 2.1 mmol, 86%).

2-Hydroxy-6-(4-methanesulfonyl-piperazin-1-yl)-naphthalene-1-carbaldehyde4-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (150 mg, 0.6 mmol),1-methanesulfonyl-piperazine hydrochloride 4b (132 mg, 0.66 mmol),sodium-tert-butoxide (190 mg, 1.98 mmol),tris-(dibenzylideneacetone)dipalladium(0) (38 mg, 41 μmol),(2-biphenyl)di-tert-butylphosphine (27 mg, 90 μmol) were dissolved in 18mL of dry dioxane. The resulted suspension was heated to 100° C. for 3h. The reaction mixture was evaporated and partitioned between 30 mL ofdichloromethane and 30 mL of water. The pH of the aqueous phase wasadjusted to neutral with acetic acid then was separated, and extractedwith another 30 mL portion of dichloromethane. The combined organicphases were dried over sodium sulfate, filtered and evaporated. Theresulted solid material was purified by chromatography with chloroformas eluent. The obtained crude product was triturated with diethyl etherto afford 4-1 as a dark yellow solid (78 mg, 0.23 mmol, 39%).

LC/MS ESI: M+H=335, Rt: 3.42 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.68(br. s., 1H), 10.76 (s, 1H), 8.80 (d, J=9.3 Hz, 1H), 7.98 (d, J=9.0 Hz,1H), 7.48 (dd, J=9.3, 2.8 Hz, 1H), 7.27 (d, J=2.8 Hz, 1H), 7.16 (d,J=9.0 Hz, 1H), 3.31 (br. s., 8H), 2.94 (s, 3H).

The following compounds were made by the above procedure:

No. MW M + H Rt 4-1

334.4 335 3.42 4-2

362.4 363 2.76 4-3

348.4 349 3.05 4-4

348.4 349 3.35 4-5

362.4 363 3.62

Example 5 Synthesis ofN-[1-(5-Formyl-6-hydroxy-naphthalen-2-yl)-pyrrolidin-3-yl]-N-methyl-acetamide5-2

2-Hydroxy-6-(3-methylamino-pyrrolidin-1-yl)-naphthalene-1-carbaldehydedihydrochloride 5-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (150 mg, 0.6 mmol),methyl-pyrrolidin-3-yl-carbamic acid tert-butyl ester (144 mg, 0.72mmol), sodium-tert-butoxide (253 mg, 2.64 mmol),tris-(dibenzylideneacetone)dipalladium(0) (38 mg, 42 μmol),(2-biphenyl)di-tert-butylphosphine (25 mg, 90 μmol) were dissolved in 16mL of dry dioxane. The resulting tan slurry was heated to 100° C. for 3h. The reaction mixture was evaporated and partitioned between 20 mL ofchloroform and 20 mL of water. The pH of the aqueous phase was adjustedto neutral with acetic acid then was separated, and extracted withanother 20 mL portion of chloroform. The combined organic phases weredried over sodium sulfate, filtered and evaporated. The resulting solidmaterial was purified by chromatography with chloroform as eluent. Theobtained crude intermediate was triturated with diethyl ether. Theresulted solid was dissolved in ethyl acetate containing HCl (10 mL) at0° C. and let to reach room temperature. After 2 h, the resultingsuspension was filtered and washed with diethyl ether to obtain 5-1 (78mg, 0.23 mmol, 99%).

LC/MS ESI: M+H=271, Rt: 2.67 min; ¹H NMR (400 MHz, DMSO-d₆) salt δ ppm11.67 (br. s., 1H), 10.75 (s, 1H), 9.37 (br. s., 2H), 8.80 (d, J=9.3 Hz,1H), 7.95 (d, J=9.0 Hz, 1H), 7.08-7.22 (m, 2H), 6.90 (d, J=2.5 Hz, 1H),3.84-3.93 (m, 1H), 3.59-3.67 (m, 1H), 3.51-3.59 (m, 2H), 3.28-3.41 (m,1H), 2.62 (t, J=5.4 Hz, 3H), 2.32-2.45 (m, 1H), 2.14-2.30 (m, 1H).

N-[1-(5-Formyl-6-hydroxy-naphthalen-2-yl)-pyrrolidin-3-yl]-N-methyl-acetamide5-2

2-Hydroxy-6-(3-methylamino-pyrrolidin-1-yl)-naphthalene-1-carbaldehydedihydrochloride 5-1 (60 mg, 018 mmol) was dissolved in 3 mL of abs.dichloromethane and acetic anhydride (54 mg, 53 mmol) was added. After30 min stirring at room temperature 1 mL of saturated sodium bicarbonatewas added. The mixture was transferred into a reparatory funnel, and theorganic layer was separated, dried over sodium sulfate, evaporated andtriturated with diethyl ether. The resulting slurry was filtered off anddried to afford 5-2 (40 mg, 13 mmol, 73%).

LC/MS ESI: M+H=313, Rt: 3.46 min; ¹H NMR (400 MHz, CDCl₃) rotamers A andB in a ratio of 70:30 δ ppm 12.81 (s, 1H, A+B), 10.75 (s, 1H, A+B),8.17-8.27 (m, 1H, A+B), 7.76-7.87 (m, 1H, A+B), 6.97-7.13 (m, 2H, A+B),6.78 (br. s., 1H, A+B), 5.37-5.55 (m, 0.7H, A), 4.57-4.79 (m, 0.3H, B),3.50-3.68 (m, 2H, A+B), 3.24-3.46 (m, 2H, A+B), 2.98 (s, 2.1H, A), 2.92(s, 0.9H, B), 2.24-2.37 (m, 0.7H, A), 2.23 (s, 0.9H, B), 2.14 (s, 2.1H,A), 2.04-2.13 (m, 0.7H, A).

Example 6 Synthesis ofN-[1-(5-Formyl-6-hydroxy-naphthalen-2-yl)-pyrrolidin-3-yl]-acetamide(IRE-1508) 6-1

3-Acetylamino-pyrrolidine-1-carboxylic acid tert-butyl ester 6a

3-Amino-pyrrolidine-1-carboxylic acid tert-butyl ester (2.0 g, 10.75mmol) was dissolved in 20 mL of dichloroethane and acetic anhydride(1.15 g, 11.29 mmol) and triethylamine (1.14 g, 11.29) was added. Afterstirring for 1 h at room temperature the mixture was evaporated and theresidue was pushed through a plug of silica with chloroform as eluent toafford 6a (2.2 g, 9.65 mmol, 90%).

N-Pyrrolidin-3-yl-acetamide hydrochloride 6b

3-Acetylamino-pyrrolidine-1-carboxylic acid tert-butyl ester 6a (2.2 g,9.65 mmol) was dissolved in ethyl acetate containing HCl (20 mL) at 0°C. and let to reach room temperature. After 2 h stirring, the suspensionwas evaporated (hygroscopic if filtered). Ethanol and then diethyl etherwas evaporated from the oily crude material to remove HCl and afford 6b(1.15 g, 7.02 mmol, 73%) as a brown oil.

N-[1-(5-Formyl-6-hydroxy-naphthalen-2-yl)-pyrrolidin-3-yl]-acetamide 6-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (151 mg, 0.6 mmol),N-pyrrolidin-3-yl-acetamide hydrochloride 6b (115 mg, 0.90 mmol),sodium-tert-butoxide (280 mg, 3.0 mmol),tris-(dibenzylideneacetone)dipalladium(0) (38 mg, 42 μmol),(2-biphenyl)di-tert-butylphosphine (27 mg, 90 μmol) were dissolved in 12mL of dry dioxane. The resulting tan slurry was heated to 100° C. for 3h. The reaction mixture was evaporated and partitioned between 20 mL ofchloroform and 20 mL of water. The pH of the aqueous phase was adjustedto neutral with acetic acid then was separated, and extracted withanother 20 mL portion of chloroform. The combined organic phases weredried over sodium sulfate, filtered and evaporated. The resulting solidmaterial was purified by eluting with 98:2 chloroform/methanol. Theobtained crude product was triturated with diethyl ether to afford 6-1.(34 mg, 0.11 mmol, 19%).

LC/MS ESI: M+H=299, Rt: 3.22 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.55(s, 1H), 10.75 (s, 1H), 8.75 (d, J=9.3 Hz, 1H), 8.17 (d, J=7.0 Hz, 1H),7.92 (d, J=9.0 Hz, 1H), 7.06-7.15 (m, 2H), 6.82 (d, J=2.5 Hz, 1H),4.34-4.44 (m, 1H), 3.56 (dd, J=9.7, 6.4 Hz, 1H), 3.41-3.50 (m, 1H),3.34-3.40 (m, 1H), 3.14 (dd, J=9.8, 4.3 Hz, 1H), 2.15-2.26 (m, 1H),1.86-1.96 (m, 1H), 1.82 (s, 3H).

The following compounds were made by the above procedure:

No. MW M + H Rt 6-1

189.34 299 3.22 6-2

396.5  397 2.73

Example 7 Synthesis of trifluoromethanesulfonic acid8-formyl-7-hydroxy-naphthalen-2-yl ester 7-1

Trifluoromethanesulfonic acid 7-hydroxy-naphthalen-2-yl ester 7a

Pyridine (2.85 mL, 2.8 g, 35 mmol, dried over KOH) was added to asuspension of 2,7-dihydroxynaphthalene (0.8 g, 5 mmol) indichloromethane (10 mL, distilled from CaH₂). The reaction mixture wascooled in ice water, trifluoromethanesulfonic anhydride (1 mL, 1.64 g, 6mmol) was added dropwise below 5° C. and the mixture was stirred in thecooling bath for 2 h. 1 N Hydrochloric acid (12 mL) was then added; theaqueous layer was separated and extracted with dichloromethane (2×5 mL).The combined organic layers were washed with water (3×5 mL), dried overmagnesium sulfate and evaporated to dryness. The oily crude product waspurified by column chromatography on silica gel eluting with 2:1hexane/ethyl acetate. In this manner7-trifluoromethanesulfonyoxy-2-naphthol 7a (0.70 g, yield: 48%) wasobtained as a thick oil which solidified upon standing.

LC/MS ESI: M−H=291, Rt: 3.83 min

This intermediate was used in the next step without furtherpurification.

Synthesis of trifluoromethanesulfonic acid8-formyl-7-hydroxy-naphthalen-2-yl ester 7-1

To a solution of 7-trifluoromethanesulfonyoxy-2-naphthol 7a (0.40 g,1.37 mmol) in dichloromethane (10 mL, distilled from CaH₂) stirred in anice water bath, titanium tetrachloride (0.30 mL, 0.52 g, 2.74 mmol) andthen dichloromethyl methyl ether (0.37 mL, 0.47 g, 4.1 mmol) were addedbelow 10° C. The mixture was stirred in the cooling bath for 2 h. 2 NHydrochloric acid (10 mL) was then added; the aqueous layer wasseparated and extracted with dichloromethane (2×5 mL). The combinedorganic layers were washed with brine (5×5 mL), dried over magnesiumsulfate and evaporated to dryness. The crude product was purified bycolumn chromatography on silica gel eluting with 4:1 hexane/diethylether. In this manner 7-1 (0.26 g, yield: 59%) was obtained as asemisolid. Trituration of a sample with diisopropyl ether yielded awhite powder.

LC/MS ESI: M+H=319, Rt: 4.09 min; ¹H NMR (400 MHz, CDCl₃) δ ppm 13.22(s, 1H), 10.73 (s, 1H), 8.22 (d, J=2.3 Hz, 1H), 8.03 (d, J=9.3 Hz, 1H),7.92 (d, J=8.8 Hz, 1H), 7.36 (dd, J=9.0, 2.3 Hz, 1H), 7.25 (d, J=9.0 Hz,1H).

Example 8 Synthesis of (5-formyl-6-hydroxy-naphthalen-2-yloxy)-aceticacid 8-1

6-hydroxy-naphthalen-2-yloxy)-acetic acid ethyl ester 8a

To a solution of 2,6-dihydroxynaphthalene (3 g, 18.75 mmol) indimethylformamide (90 mL) NaH (822 mg, ˜60% oil dispersion) was added.After 1 h stirring bromoacetic acid ethyl ester (2.29 mL, 20.62 mmol)was added. The suspension was stirred for another 4 h at roomtemperature. The solvent was evaporated under reduced pressure, thensuspended in water (200 mL) and acidified with 10% hydrochloric acid,then extracted with ethyl acetate (2×150 mL). The combined organiclayers were dried over magnesium sulfate and evaporated to dryness. Thecrude product was purified by column chromatography on silica gel,eluting with chloroform to afford 8a as a solid (1.54 g, 6.26 mmol,33%).

LC/MS ESI: M+H=247, Rt: 3.28 min.

This intermediate was used in the next step without furtherpurification.

(5-formyl-6-hydroxy-naphthalen-2-yloxy)-acetic acid ethyl ester 8b

6-Hydroxy-naphthalen-2-yloxy)-acetic acid ethyl ester 8a (2.11 g, 8.58mmol) in dichloromethane (45 mL, distilled from CaH₂) was added to astirred solution of titanium tetrachloride (1.55 mL, 14.3 mmol) anddichloromethyl methyl ether (2.3 mL, 25.7 mmol) in dichloromethane (35mL, distilled from CaH₂) at 0° C., and the mixture was stirred in thecooling bath for 1 h, then at room temperature overnight. 1 Nhydrochloric acid (80 mL) was then added; the organic layer wasseparated and extracted with 1 N hydrochloric acid (2×80 mL), then with100 mL aqueous EDTA disodium salt. The organic layer was washed with 50mL of saturated sodium bicarbonate, dried over magnesium sulfate andevaporated to dryness. The crude product was purified by columnchromatography on silica gel, eluting with hexane/ethyl acetate toafford 8b as a reddish solid (355 mg, 1.29 mmol, 15%).

LC/MS (ESI): M+H=275, Rt: 3.66 min.

(5-formyl-6-hydroxy-naphthalen-2-yloxy)-acetic acid 8-1

(5-Formyl-6-hydroxy-naphthalen-2-yloxy)-acetic acid ethyl ester 8b (340mg, 1.24 mmol) was dissolved in 40 mL 1:1 mixture of dioxane-10% aq.sodium hydroxide and stirred for 30 min at room temperature. 50 mLDichloromethane was added to the reaction mixture, the aqueous layer wasseparated and washed with 50 mL dichloromethane, acidified with 1Nhydrochloric acid and the precipitate was filtered off and washed withwater to afford 8-1 as a pink solid (270 mg, 1.09 mmol, 88%).

LC/MS ESI: M−H=245, Rt: 2.99 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.69(br. s., 1H), 10.76 (s, 1H), 8.86 (m, 1H), 8.06 (d, J=8.8 Hz, 1H), 7.30(m, 2H), 7.21 (d, J=9.2 Hz, 1H), 4.76 (s, 2H).

Example 9 Synthesis of2-Hydroxy-7-(2-morpholin-4-yl-2-oxo-ethoxy)-naphthalene-1-carbaldehyde9-1

8-Formyl-7-hydroxy-naphthalen-2-yloxy)-acetic acid (123 mg, 0.5 mmol),1-hydroxybenzotriazole (149 mg, 1.1 mmol), morpholine (95 μL, 1.1 mmol),triethylamine (350 μL, 2.5 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (210 mg, 1.1 mmol) were dissolved intetrahydrofuran (4 mL), and the mixture was stirred overnight at roomtemperature. The solvent was evaporated under reduced pressure and tothe residue was poured 30 mL saturated sodium bicarbonate and extractedwith chloroform (2×30 mL). The combined organic layers were dried overmagnesium sulfate and evaporated to dryness. The crude product waspurified by column chromatography on silica gel, eluting withchloroform, then was finally triturated with diethyl ether yielding 9-1as a yellow powder (65 mg, 0.206 mmol, 41%).

LC/MS ESI: M+H=316, Rt: 3.11 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.90(br. s., 1H), 10.77 (s, 1H), 8.38 (d, J=2.5 Hz, 1H), 8.04 (d, J=8.8 Hz,1H), 7.81 (d, J=9.0 Hz, 1H), 7.10 (dd, J=8.9, 2.6 Hz, 1H), 7.05 (d,J=9.0 Hz, 1H), 4.97 (s, 2H), 3.69 (br. s., 2H), 3.60 (br. s., 2H), 3.54(br. s., 2H), 3.47 (br. s., 2H).

The following compounds were made by the above procedure:

No. MW M + H Rt 9-1

315.3 316 3.11 9-2

342.4 343 2.52 9-3

358.4 359 2.50 9-4

316.4 317 2.49 9-5

328.3 329 2.99 9-6

315.3 316 3.07

Example 10 Synthesis of2-Hydroxy-7-[2-(4-methyl-piperazin-1-yl)-2-oxo-ethoxy]-naphthalene-1-carbaldehyde10-1

(8-Formyl-7-hydroxy-naphthalen-2-yloxy)-acetyl chloride 10a

Thionyl chloride (20 mL) was added to the8-formyl-7-hydroxy-naphthalen-2-yloxy)-acetic acid (200 mg, 0.81 mmol),and the mixture was refluxed for 2 h. The solvent was evaporated underreduced pressure and the residue was used in the next step withoutfurther purification.

2-Hydroxy-7-[2-(4-methyl-piperazin-1-yl)-2-oxo-ethoxy]-naphthalene-1-carbaldehyde10-1

To a solution of N-methylpiperazine (123 μL, 0.89 mmol) andtriethylamine (340 μL, 2.44 mmol) in 15 mL dichloroethane at 0° C.,8-Formyl-7-hydroxy-naphthalen-2-yloxy)-acetyl chloride 10a was added andthe mixture was allowed to warm to room temperature. Then the mixturewas extracted with water (25 mL) and the aqueous layer was washed withdichloromethane (2×25 mL). The combined organic layers were dried overmagnesium sulfate and evaporated to dryness. The crude product waspurified by column chromatography on silica gel, eluting with 40:1chloroform/methanol. The obtained product was triturated with diethylether and filtered to give 10-1 (20 mg, 60.9 μmol, 8%).

LC/MS ESI: M+H=329, Rt: 2.49 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.92(br. s., 1H), 10.77 (s, 1H), 8.37 (d, J=2.5 Hz, 1H), 8.04 (d, J=9.0 Hz,1H), 7.80 (d, J=9.0 Hz, 1H), 7.10 (dd, J=8.8, 2.5 Hz, 1H), 7.05 (d,J=8.8 Hz, 1H), 4.95 (s, 2H), 3.44-3.56 (m, 4H), 2.43 (br. s., 2H), 2.33(br. s., 2H), 2.23 (s, 3H).

Example 11 Synthesis of2-Hydroxy-6-[4-(4-methyl-piperazine-1-carbonyl)-phenoxy]-naphthalene-1-carbaldehyde11-1

4-(6-Hydroxy-naphthalen-2-yloxy)-benzonitrile 11a

Naphthalene-2,6-diol (2.28 g, 14.25 mmol), 4-fluoro-benzonitrile (1.72g, 14.25 mmol) and K₂CO₃ (1.96 g, 14.25 mmol) were dissolved in 60 mL ofDMF and the mixture was heated to 150° C. for 2 h. The reaction mixturewas partitioned between water and dichloromethane. The organic layer wasseparated and extracted, washed with 1N hydrochloric acid, separated,dried over sodium sulfate, filtered and evaporated. The obtained crudeproduct was purified by column chromatography on silica gel, elutingwith 20:1 chloroform/methanol to afford 11a (610 mg, 2.34 mmol, 16%).

4-(5-Formyl-6-hydroxy-naphthalen-2-yloxy)-benzonitrile 11b

4-(6-Hydroxy-naphthalen-2-yloxy)-benzonitrile 11a (550 mg, 2.03 mmol) indichloromethane (10 mL, distilled from CaH₂) was added to a stirredsolution of titanium tetrachloride (0.67 mL, 3.39 mmol) anddichloromethyl methyl ether (0.62 mL, 6.09 mmol) in dichloromethane (10mL, distilled from CaH₂) at 0° C., and the mixture was stirred at 0° C.for 1 h, then at room temperature overnight. 1 N hydrochloric acid (20mL) was then added; the organic layer was separated and extracted with 1N hydrochloric acid (2×20 mL). The organic layer was washed with 10 mLof saturated sodium bicarbonate, dried over magnesium sulfate andevaporated to dryness. The crude product was purified by columnchromatography on silica gel, eluting with dichloromethane to afford 11b(190 mg, 0.66 mmol, 32%).

LC/MS ESI: M−H=288, Rt: 4.05 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.80(br. s., 1H), 10.79 (s, 1H), 9.06 (d, J=9.2 Hz, 1H), 8.10 (d, J=8.8 Hz,1H), 7.84 (m, 2H), 7.65 (d, J=2.4 Hz, 1H), 7.45 (dd, J=9.2, 2.8 Hz, 1H)7.27 (d, J=8.8 Hz, 1H), 7.07 (m, 2H).

4-(5-Formyl-6-hydroxy-naphthalen-2-yloxy)-benzoic acid 11c

4-(5-Formyl-6-hydroxy-naphthalen-2-yloxy)-benzonitrile 11b (170 mg, 0.59mmol) was dissolved in a mixture of 20 mL of methanol and 20 mL of 10%aqueous sodium hydroxide. The reaction was heated to 80° C. for 12 h.The cooled reaction mixture was acidified with conc. aqueoushydrochloric acid and the resulting precipitate was filtered and thecrude product was purified by column chromatography on silica gel,eluting with 20:1 chloroform/methanol, to afford 11-1 (60 mg, 0.19 mmol,32%).

LC/MS ESI: M−H=307, Rt: 3.69 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm, 10.79(s, 1H), 9.04 (d, J=9.6 Hz, 1H), 8.09 (d, J=9.2 Hz, 1H), 7.95 (m, 2H),7.61 (d, J=2.4 Hz, 1H), 7.45 (dd, J=9.2, 2.8 Hz, 1H), 7.27 (d, J=8.8 Hz,1H), 7.07 (m, 2H).

2-Hydroxy-6-[4-(4-methyl-piperazine-1-carbonyl)-phenoxy]-naphthalene-1-carbaldehyde11-1

4-(5-Formyl-6-hydroxy-naphthalen-2-yloxy)-benzoic acid 11c (40 mg, 0.13mmol), 1-hydroxybenzotriazole (38 mg, 0.29 mmol), N-methyl-piperazine(32 μL, 0.39 mmol), triethylamine (90 μL, 0.65 mmol) and1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (55 mg,0.29 mmol) were dissolved in dimethylformamide (4 mL), and the mixturewas stirred overnight at room temperature. The solvent was evaporatedunder reduced pressure and to the residue was poured 10 mL saturatedsodium bicarbonate and extracted with chloroform (2×10 mL). The combinedorganic layers were dried over magnesium sulfate and evaporated todryness. The crude product 11-1 was purified by column chromatography onsilica gel, eluting with chloroform, then was finally isolated as a HClsalt after treatment with ethyl acetate containing HCl (29 mg, 0.07mmol, 54%).

LC/MS ESI: M+H=391, Rt: 2.89 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm, 11.81(s, 1H), 10.90 (br, s, 1H), 10.79 (s, 1H), 9.05 (d, J=9.2 Hz, 1H), 8.08(d, J=8.8 Hz, 1H), 7.56 (d, J=2.8 Hz, 1H), 7.49 (d, J=8.8 Hz, 1H), 7.43(dd, J=9.2, 2.4 Hz, 1H), 7.30 (d, J=8.8 Hz, 1H), 7.08 (d, J=8.8 Hz, 2H),4.20 (br, 2H) 3.40 (br, 4H) 3.05 (br, 2H) 2.77 (s, 3H).

Example 12 Synthesis of2-Hydroxy-6-[4-methyl-5-(morpholine-4-carbonyl)-thiazol-2-yl]-naphthalene-1-carbaldehyde12-1

2-Bromo-4-methyl-thiazole-5-carbonyl chloride 12a

2-Bromo-4-methyl-thiazole-5-carboxylic acid (250 mg, 1.13 mmol) wasdissolved in 5 mL of thionyl chloride. After refluxing for 1 h themixture was evaporated, dissolved in 10 mL of toluene and evaporatedagain to afford 2-bromo-4-methyl-thiazole-5-carbonyl chloride 12a. (228mg, 0.91 mmol, 84%).

(2-Bromo-4-methyl-thiazol-5-yl)-morpholin-4-yl-methanone 12b

To a stirred mixture of morpholine (87 mg, 1.0 mmol) anddiisopropyl-ethyl-amine (184 mg, 1.43 mmol) in 7 mL of abs.dichloroethane at 0° C., 2-bromo-4-methyl-thiazole-5-carbonyl chloride12a (228 mg, 0.95 mmol) in 7 mL of abs. dichloroethane was addeddropwise. The mixture was stirred for an additional 2 h at roomtemperature. The reaction mixture was extracted with 15 mL of saturatedsodium bicarbonate; the organic layer was separated, dried over sodiumsulfate, filtered and evaporated to afford(2-bromo-4-methyl-thiazol-5-yl)-morpholin-4-yl-methanone 12b as a yellowoil. (226 mg, 78 mmol, 82%).

2-Hydroxy-6-[4-methyl-5-(morpholine-4-carbonyl)-thiazol-2-yl]-naphthalene-1-carbaldehyde12-1

(2-bromo-4-methyl-thiazol-5-yl)-morpholin-4-yl-methanone 12b (226 mg,0.78 mmol),2-Hydroxy-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-naphthalene-1-carbaldehyde(12c, WO2008154484) (231 mg, 0.78 mmol), sodium carbonate (660 mg, 6.24mmol), and tetrakis(triphenylphosphine)palladium (27 mg, 0.023 mmol)were dissolved in a mixture of 18 mL DMF and 18 mL water. The reactionmixture was stirred at 120° C. under argon for 1 h. The reaction mixturewas evaporated to dryness and the solid residue was partitioned betweenchloroform and water. The aqueous phase was acidified with acetic acidto pH 6. The organic phase was separated, and the aqueous layer wasextracted once more with chloroform. The combined organic phases weredried over sodium sulfate, filtered and evaporated. The residue waspurified by column chromatography with chloroform as eluent. The crudeproduct was triturated with diethyl ether, filtered off and air dried,affording 12-1 (116 mg, 0.31 mmol, 39%).

LC/MS ESI: M+H=383, Rt: 3.47 min; ¹H NMR (400 MHz, CDCl₃) δ ppm 13.21(s, 1H), 10.83 (s, 1H), 8.41 (d, J=9.0 Hz, 1H), 8.36 (d, J=1.5 Hz, 1H),8.13 (dd, J=8.9, 1.5 Hz, 1H), 8.06 (d, J=9.3 Hz, 1H), 7.21 (d, J=9.0 Hz,1H), 3.73-3.78 (m, 4H), 3.65-3.73 (m, 4H), 2.55 (s, 3H).

The following compounds were made by the above procedure.

No. MW M + H 12-1

382.4 383 12-2

381.5 382 12-3

381.5 382 12-4

395.5 396 12-5

365.4 366 12-6

380.5 381 12-7

404.5 405 12-8

394.4 395 12-9

368.5 369 12-10

382.5 3.83 12-11

350.4 351 12-12

405.5 406 12-13

366.4 367 12-14

370.4 371 12-15

425.5 426 12-16

345.2 346 12-17

347.1 348 12-18

345.2 346 12-19

374.1 375 12-20

346.1 347

Example 13 Synthesis of2-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiazole-5-carboxylicacid (1-methyl-piperidin-4-yl)-amide 13-1

4-[(2-Bromo-4-methyl-thiazole-5-carbonyl)-amino]piperidine-1-carboxylicacid tert-butyl ester 13a

To a stirred mixture of 4-amino-piperidine-1-carboxylic acid tert-butylester (801 mg, 4.0 mmol) and diisopropyl-ethyl-amine (517 mg, 4.0 mmol)in 40 mL of abs. dichloromethane at 0° C.,2-bromo-4-methyl-thiazole-5-carbonyl chloride 12a (960 mg, 4.0 mmol) in10 mL of abs. dichloroethane was added dropwise. The mixture was stirredfor an additional 2 h at room temperature. The reaction mixture wasextracted with 50 mL of saturated sodium bicarbonate; the organic layerwas separated, dried over sodium sulfate, filtered and evaporated toafford 13a (1.1 g, 2.72 mmol, 68%).

2-Bromo-4-methyl-thiazole-5-carboxylic acid piperidin-4-ylamidehydrochloride 13b

4-[(2-Bromo-4-methyl-thiazole-5-carbonyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester 13a (660 mg, 1.63 mmol) was suspended in cca. 4MHCl in ethyl acetate (20 mL) at ° C. and let to warm up to roomtemperature. After 3 h stirring, the suspension was evaporated andfiltered with diethyl ether to obtain 13b (276 mg, 0.81 mmol, 50%).

2-Bromo-4-methyl-thiazole-5-carboxylic acid(1-methyl-piperidin-4-yl)-amide 13c

To a solution of 2-bromo-4-methyl-thiazole-5-carboxylic acidpiperidin-4-ylamide hydrochloride 13b (457 mg, 1.34 mmol) in methanol (5mL), sodium bicarbonate (124 mg, 1.48 mmol), 37% aqueous formaldehyde(1.091 g, 13.5 mmol) and NaBH₃CN (101 mg, 1.6 mmol) were added. Thereaction was stirred overnight at room temperature, then evaporated. Theresidue was suspended in saturated sodium bicarbonate and extracted withethyl acetate. The combined organic phase were dried over sodiumsulfate, filtered and evaporated. The residue was purified by columnchromatography with chloroform:methanol as eluent, to afford 13c as asolid (250 mg, 0.78 mmol, 58%).

2-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiazole-5-carboxylicacid (1-methyl-piperidin-4-yl)-amide 13-1

2-hydroxy-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-naphthalene-1-carbaldehyde(12c, 149 mg, 0.5 mmol), 2-bromo-4-methyl-thiazole-5-carboxylic acid(1-methyl-piperidin-4-yl)-amide 13c (159 mg, 0.5 mmol), sodium carbonate(318 mg, 3 mmol), and tetrakis(triphenylphosphine)palladium (17 mg,0.015 mmol) were dissolved in a mixture of 5 mL DMF and 5 mL water. Thereaction mixture was stirred at 100° C. under argon for 1 h. Thereaction mixture was evaporated to dryness and the solid residue waspartitioned between dichloromethane and water. The organic phase wasseparated, and the aqueous layer was extracted once more withchloroform. The combined organic phases were dried over sodium sulfate,filtered and evaporated. The residue was purified by columnchromatography with chloroform:methanol as eluent. The crude product wastriturated with diethyl ether, filtered off and air dried, affording13-1 (45 mg, 0.11 mmol, 22%).

LC/MS ESI: M+H=410, Rt: 2.79 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.70(s, 1H), 9.09 (d, J=9.0 Hz, 1H), 8.38 (d, J=1.8 Hz, 1H), 8.19 (d, J=7.5Hz, 1H), 8.13 (d, J=9.3 Hz, 1H), 8.02 (dd, J=8.9, 1.9 Hz, 1H), 7.18 (d,J=9.0 Hz, 1H), 3.72-3.82 (m, 1H), 2.86-2.97 (m, 2H), 2.61 (s, 3H), 2.31(s, 3H), 2.20-2.29 (m, 2H), 1.79-1.89 (m, 2H), 1.62-1.72 (m, 2H).

The following compounds were made by the above procedure.

No. MW M + H Rt 13-1

409.5 410 2.80 13-2

408.5 409 2.81 13-3

408.5 409 2.78 13-4

394.5 395 2.76 13-5

388.5 389 2.91

Example 14 Synthesis of2-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiazole-5-carboxylicacid piperidin-4-ylamide hydrochloride 14-1

4-[(2-Bromo-4-methyl-thiazole-5-carbonyl)-amino]piperidine-1-carboxylicacid tert-butyl ester 14a

To a stirred mixture of 4-amino-piperidine-1-carboxylic acid tert-butylester (801 mg, 4.0 mmol) and diisopropyl-ethyl-amine (517 mg, 4.0 mmol)in 40 mL of abs. dichloromethane at 0° C.,2-bromo-4-methyl-thiazole-5-carbonyl chloride (see Example “Arev”/stepA;960 mg, 4.0 mmol) in 10 mL of abs. dichloroethane was added dropwise.The mixture was stirred for an additional 2 h at room temperature. Thereaction mixture was extracted with 50 mL of saturated sodiumbicarbonate; the organic layer was separated, dried over sodium sulfate,filtered and evaporated to afford 14a. (1.1 g, 2.72 mmol, 68%).

2-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiazole-5-carboxylicacid piperidin-4-ylamide hydrochloride 14-1

2-Hydroxy-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-naphthalene-1-carbaldehyde12c (298 mg, 1 mmol),4-[(2-Bromo-4-methyl-thiazole-5-carbonyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester 14a (404 mg, 1 mmol), sodium carbonate (636 mg, 6mmol), and tetrakis(triphenylphosphine)palladium (34 mg, 0.03 mmol) weredissolved in a mixture of 9 mL DMF and 9 mL water. The reaction mixturewas stirred at 100° C. under argon for 1 h. The reaction mixture wasevaporated to dryness and the solid residue was partitioned betweenchloroform and brine. The organic phase was separated, and the aqueouslayer was extracted once more with chloroform. The combined organicphases were dried over sodium sulfate, filtered and evaporated. Theresidue was purified by column chromatography with chloroform as eluent.The crude product was triturated with diethyl ether, filtered off andair dried. The resulted solid was dissolved in 5 mL methanol, and ethylacetate containing HCl (2 mL) was added at 0° C. and allowed to reachroom temperature. After 3 h stirring, the suspension concentrated underreduced pressure and triturated with diethyl ether to obtain 14-1 (163mg, 0.38 mmol, 38%).

LC/MS ESI: M+H=396, Rt: 2.80 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.09(br. s., 1H), 10.79 (s, 1H), 9.09 (d, J=9.0 Hz, 1H), 8.86-9.06 (m, 2H),8.50 (d, J=2.0 Hz, 1H), 8.46 (d, J=7.5 Hz, 1H), 8.29 (d, J=9.0 Hz, 1H),8.11 (dd, J=9.0, 2.0 Hz, 1H), 7.40 (d, J=9.0 Hz, 1H), 3.96-4.10 (m, 1H),3.29 (d, J=12.8 Hz, 2H), 2.91-3.07 (m, 2H), 2.63 (s, 3H), 1.91-2.07 (m,2H), 1.72-1.87 (m, 2H).

The following compounds were made by the above procedure:

No. MW M + H Rt 14-1

395.5 396 2.80 14-2

394.5 395 2.73 14-3

394.5 395 2.00 14-4

394.5 395 1.98 14-5

428.5 429 1.98 14-6

380.5 381 2.89 14-7

380.5 381 2.68 14-8

374.4 375.2 2.90 14-9

374.4 375 2.71 14-10

364.4 365 2.67 14-11

366.4 367 2.63 14-12

378.4 379 2.69 14-13

375.4 376 2.82

Example 15 Synthesis of2-Hydroxy-6-(3-morpholin-4-yl-3-oxo-propenyl)-naphthalene-1-carbaldehyde15-3

3-(5-formyl-6-hydroxy-naphthalen-2-yl)-acrylic acid ethyl ester 15-1

To a solution of 6-bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (1 g, 4mmol) in 4 mL dimethylformamide, ethyl acrylate (521 μL, 4.8 mmol),triethylamine (780 μL, 5.6 mmol) andtetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol) were added andthe mixture was stirred under nitrogen at 100° C. for 1 h. The solventwas evaporated under reduced pressure, then the residue was suspended inwater (500 mL) and extracted with dichloromethane (2×50 mL). Thecombined organic layers were dried over magnesium sulfate and evaporatedto dryness. The crude product was purified by column chromatography onsilica gel, eluting with toluene to afford 15-1 as a yellow solid (600mg, 2.22 mmol, 55%).

LC/MS ESI: M+H=271, Rt: 3.18 min; ¹H NMR (400 MHz, CDCl₃) δ ppm 13.17(s, 1H), 10.81 (s, 1H), 8.36 (d, J=8.8 Hz, 1H), 8.00 (d, J=9.0 Hz, 1H),7.89 (s, 1H), 7.74-7.84 (m, 2H), 7.18 (d, J=9.0 Hz, 1H), 6.54 (d, J=15.8Hz, 1H), 4.30 (q, J=7.0 Hz, 2H), 1.36 (t, J=7.2 Hz, 3H).

3-(5-Formyl-6-hydroxy-naphthalen-2-yl)-acrylic acid 15-2

3-(5-Formyl-6-hydroxy-naphthalen-2-yl)-acrylic acid ethyl ester 15-1(534 mg; 1.97 mmol) was dissolved in a mixture 25 mL of dioxane and 20mL of 1N sodium hydroxide and was heated to 50° C. for 0.5 h. Thereaction mixture was extracted with 30 mL of chloroform and the aqueouslayer was cooled to 0° C. and 6N hydrochloric acid was added dropwise.The precipitated solid was filtered, washed with distilled water toafford 15-2. (418 mg, 1.55 mmol, 87%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.39 (br. s., 1H), 12.04 (s, 1H), 10.79(s, 1H), 8.95 (d, J=9.0 Hz, 1H), 8.10-8.22 (m, 2H), 7.96 (dd, J=8.9, 1.6Hz, 1H), 7.69 (d, J=16.1 Hz, 1H), 7.29 (d, J=9.0 Hz, 1H), 6.63 (d,J=15.8 Hz, 1H).

2-Hydroxy-6-(3-morpholin-4-yl-3-oxo-propenyl)-naphthalene-1-carbaldehyde15-3

3-(5-Formyl-6-hydroxy-naphthalen-2-yl)-acrylic acid 15-2 (73 mg, 0.3mmol), 1-hydroxybenzotriazole (89 mg, 0.66 mmol), morpholine (58 mg,0.66 mmol) and triethylamine (151 mg, 1.5 mmol) were dissolved in 5 mLTHF. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (127mg, 0.66 mmol) was added with stirring at room temperature. After 2 h, 5mL of 2N aqueous hydrochloric acid was added and the mixture was stirredfor an additional 2 h. The reaction mixture was evaporated to drynessand the solid residue was partitioned between 15 mL of chloroform and 15mL of saturated sodium bicarbonate. The aqueous phase was extracted withan additional 15 mL portion of chloroform; the combined organic phaseswere extracted with brine, dried over sodium sulfate, filtered off andevaporated. The solid material was purified by chromatography on silica,with chloroform as eluent to afford 15-3. (50 mg, 0.16 mmol, 54%).

¹H NMR (400 MHz, CDCl₃) δ ppm 13.14 (br. s., 1H), 10.81 (s, 1H), 8.34(d, J=8.8 Hz, 1H), 8.00 (d, J=9.0 Hz, 1H), 7.89 (s, 1H), 7.82 (d, J=15.6Hz, 1H), 7.80 (d, J=7.0 Hz, 1H), 7.18 (d, J=9.0 Hz, 1H), 6.95 (d, J=15.3Hz, 1H), 3.75 (br. s., 8H).

The following compounds were made by the above procedure.

No. NMR 15-1

¹H NMR (400 MHz, CDCl₃) δ ppm 13.17 (s, 1H), 10.81 (s, 1H), 8.36 (d, J =8.8 Hz, 1H), 8.00 (d, J = 9.0 Hz, 1H), 7.89 (s, 1H), 7.74-7.84 (m, 2H),7.18 (d, J = 9.0 Hz, 1H), 6.54 (d, J = 15.8 Hz, 1H), 4.30 (q, J = 7.0Hz, 2H), 1.36 (t, J = 7.2 Hz, 3H). 15-2

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.39 (br. s., 1H), 12.04 (s, 1H), 10.79(s, 1H), 8.95 (d, J = 9.0 Hz, 1H), 8.10-8.22 (m, 2H), 7.96 (dd, J = 8.9,1.6 Hz, 1H), 7.69 (d, J = 16.1 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H), 6.63(d, J = 15.8 Hz, 1H) 15-3

¹H NMR (400 MHz, CDCl₃) δ ppm 13.14 (br. s., 1H), 10.81 (s, 1H), 8.34(d, J = 8.8 Hz, 1H), 8.00 (d, J = 9.0 Hz, 1H), 7.89 (s, 1H), 7.82 (d, J= 15.6 Hz, 1H), 7.80 (d, J = 7.0 Hz, 1H), 7.18 (d, J = 9.0 Hz, 1H), 6.95(d, J = 15.3 Hz, 1H), 3.75 (br. s., 8H). 15-4

¹H NMR (400 MHz, CDCl₃) δ ppm 13.14 (s, 1H), 10.81 (s, 1H), 8.34 (d, J =8.8 Hz, 1H), 8.00 (d, J = 9.3 Hz, 1H), 7.89 (d, J = 1.5 Hz, 1H),7.74-7.84 (m, 2H), 7.17 (d, J = 9.0 Hz, 1H), 7.00 (d, J = 15.6 Hz, 1H),3.22 (s, 3H), 3.10 (s, 3H). 15-5

MW, 324.1, M + 1 325

2-Hydroxy-6-(3-morpholin-4-yl-3-oxo-propyl)-naphthalene-1-carbaldehyde16-3

3-(5-Formyl-6-hydroxy-naphthalen-2-yl)-acrylic acid ethyl ester (15-1,570 mg, 2.11 mmol) was dissolved in 10 mL ethyl acetate and 50 mg ofPd/C (10%) was added. The mixture was stirred at room temperature for 72h under hydrogen atmosphere. The catalyst was filtered off, and thefiltrate was evaporated under reduced pressure. The obtained tawny oilwas purified by column chromatography on silica gel, eluting with a 98:2mixture of toluene:methanol to afford 16-1 as a solid (212 mg, 0.78mmol, 37%).

LC/MS ESI: M+H=273, Rt: 3.72 min; ¹H NMR (400 MHz, CDCl₃) δ ppm 13.06(s, 1H), 10.80 (s, 1H), 8.28 (d, J=8.5 Hz, 1H), 7.92 (d, J=9.0 Hz, 1H),7.61 (s, 1H), 7.49 (dd, J=8.8, 1.5 Hz, 1H), 7.13 (d, J=9.0 Hz, 1H), 4.13(q, J=7.0 Hz, 2H), 3.10 (t, J=7.7 Hz, 2H), 2.71 (t, J=7.7 Hz, 2H), 1.23(t, J=7.2 Hz, 3H).

3-(5-Formyl-6-hydroxy-naphthalen-2-yl)-propionic acid ethyl ester 16-1(158 mg, 0.58 mmol) was added to 50 mL of a 1:1 mixture ofdioxane-sodium hydroxide (10%) and stirred for 30 min at roomtemperature. 50 mL Dichloromethane was added, the aqueous layer wasseparated and washed with 50 mL of dichloromethane. The aqueous layerwas acidified with 1N hydrochloric acid, and the precipitate wasfiltered and washed three times with water., then dried under aninfrared lamp to afford 16-2 as a solid (94 mg, 0.39 mmol, 63%).

LC/MS ESI: M+H=245, Rt: 2.28 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.14(br. s., 1H), 11.89 (br. s., 1H), 10.79 (s, 1H), 8.83 (d, J=8.8 Hz, 1H),8.06 (d, J=9.0 Hz, 1H), 7.70 (s, 1H), 7.52 (dd, J=8.9, 1.6 Hz, 1H), 7.21(d, J=9.0 Hz, 1H), 2.95 (t, J=7.5 Hz, 2H), 2.62 (t, J=7.5 Hz, 2H).

3-(5-Formyl-6-hydroxy-naphthalen-2-yl)-propionic acid 16-2 (150 mg, 0.67mmol), 1-hydroxybenzotriazole (109 mg, 0.8 mmol), morpholine (64 mg,0.74 mmol), triethylamine (270 mg, 2.68 mmol) and1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (153 mg,0.8 mmol) were dissolved in dimethylformamide (4 mL), and the mixturewas stirred overnight at room temperature. The solvent was evaporatedunder reduced pressure, 30 mL 1N hydrochloric acid was added and thesuspension was extracted with chloroform (2×30 mL), then the combinedorganic layers were washed with 30 mL saturated sodium bicarbonate. Theorganic layer was dried over magnesium sulfate and evaporated todryness. The crude product was purified by preparative HPLC to afford16-3 (26 mg, 83 μmol, 14%).

LC/MS ESI: M+H=314, Rt: 2.92 min; ¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.95(br. s., 1H), 10.79 (s, 1H), 8.83 (d, J=8.8 Hz, 1H), 8.05 (d, J=9.0 Hz,1H), 7.71 (s, 1H), 7.53 (dd, J=8.8, 1.5 Hz, 1H), 7.20 (d, J=9.0 Hz, 1H),3.45-3.53 (m, 4H), 3.39-3.45 (m, 4H), 2.95 (t, J=7.7 Hz, 2H), 2.70 (t,J=7.7 Hz, 2H).

The following compounds were made by the above procedure:

No. MW M + H Rt 16-1

272.3 273 3.75 16-2

244.3 245 2.28 16-3

313.4 314 2.92 16-4

271.3 272 2.99

Example 17 Synthesis of4-Chloro-3-(5-formyl-6-hydroxy-naphthalen-2-yl)-benzoic acid 17-1

4-Chloro-3-(5-formyl-6-hydroxy-naphthalen-2-yl)-benzoic acid 17-1

6-Bromo-2-hydroxy-naphthalene-1-carbaldehyde 1a (1.5 g, 6.0 mmol),2-chloro-5-carboxyphenylboronic acid (1.36 g, 6.6 mmol), sodiumcarbonate (660 mg, 36 mmol), and tetrakis(triphenylphosphine)palladium(200 mg, 0.17 mmol) were dissolved in a mixture of 100 mL DMF and 100 mLwater. The reaction mixture was stirred at 100° C. under argon for 6 h.The reaction mixture was extracted two times with 150 mL 1N sodiumhydroxide. The combined aqueous phases were extracted three times with50 mL of chloroform. The aqueous layer was separated, cooled to 0° C.,and was stirred vigorously, while 5N hydrochloric acid was addeddropwise. The precipitated white solid was filtered, washed with waterand diethyl ether. A 100 mg portion of the crude product was purified bychromatography on silica eluting with 95:5 chloroform/methanol to affordanalytically pure 17-1. (59 mg, 0.18 mmol, 44%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.38 (br. s., 1H), 12.04 (br. s., 1H),10.84 (s, 1H), 9.03 (d, J=9.0 Hz, 1H), 8.23 (d, J=9.0 Hz, 1H), 8.02 (d,J=2.0 Hz, 1H), 8.00 (d, J=1.8 Hz, 1H), 7.96 (dd, J=8.3, 2.0 Hz, 1H),7.75 (d, J=8.3 Hz, 1H), 7.73 (dd, J=9.0, 2.0 Hz, 1H), 7.31 (d, J=9.0 Hz,1H).

The following compounds were made by the above procedure:

No. NMR 17- 1

¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.38 (br. s., 1H), 12.04 (br. s., 1H),10.84 (s, 1H), 9.03 (d, J = 9.0 Hz, 1H), 8.23 (d, J = 9.0 Hz, 1H), 8.02(d, J = 2.0 Hz, 1H), 8.00 (d, J = 1.8 Hz, 1H), 7.96 (dd, J = 8.3, 2.0Hz, 1H), 7.75 (d, J = 8.3 Hz, 1H), 7.73 (dd, J = 9.0, 2.0 Hz, 1H), 7.31(d, J = 9.0 Hz, 1H). 17- 2

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.34 (br. s., 1H), 10.93 (s, 1H),9.20-9.31 (m, 4H), 8.22 (d, J = 8.8 Hz, 1H), 8.07 (d, J = 8.3 Hz, 1H),7.87 (dd, J = 8.4, 1.9 Hz, 1H), 7.30 (d, J = 9.0 Hz, 1H). 17- 3

¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.43 (br. s., 1H), 12.01 (br. s., 1H),10.84 (s, 1H), 9.04 (d, J = 9.0 Hz, 1H), 8.22 (d, J = 9.0 Hz, 1H), 8.06(d, J = 1.5 Hz, 1H), 8.02 (d, J = 2.0 Hz, 1H), 7.99 (dd, J = 8.0, 1.8Hz, 1H), 7.73 (dd, J = 8.8, 2.0 Hz, 1H), 7.66 (d, J = 7.8 Hz, 1H), 7.31(d, J = 9.0 Hz, 1H).

Example 18 Synthesis of5-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiophene-2-carboxylicacid 18-1

5-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiophene-2-carboxylicacid methyl ester 18a

2-Hydroxy-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-naphthalene-1-carbaldehyde12c (1.20 g, 4.0 mmol), 5-bromo-4-methyl-thiophene-2-carboxylic acidmethyl ester (1.03 g, 4.40 mmol), sodium carbonate (2.54 g, 24.0 mmol),and tetrakis(triphenylphosphine)palladium (138 mg, 0.12 mmol) weredissolved in a mixture of 100 mL DMF and 100 mL water. The reactionmixture was stirred at 105° C. under argon for 3 h. The reaction mixturewas evaporated to dryness and the solid residue was partitioned betweenchloroform and water, while the aqueous phase was acidified with aceticacid to pH 6. The organic phase was separated, and the aqueous layer wasextracted once more with chloroform. The combined organic phases weredried over sodium sulfate, filtered and evaporated. The obtained crudeproduct 18a (900 mg, 2.76 mmol, 96%) was used in the next step withoutpurification.

5-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiophene-2-carboxylicacid 18-1

5-(5-Formyl-6-hydroxy-naphthalen-2-yl)-4-methyl-thiophene-2-carboxylicacid methyl ester 18a (835 mg; 2.56 mmol) was dissolved in a mixture 30mL of dioxane and 30 mL of 1N sodium hydroxide and was stirred at 50° C.for 1 h. Charcoal was added to the mixture and was stirred for anadditional 0.5 h, then filtered. The reaction mixture was washed with 30mL of chloroform, the aqueous layer was cooled to 0° C. and 6Nhydrochloric acid was added dropwise. The precipitating solid wasfiltered, washed with distilled water to afford 18-1. (675 mg, 2.16mmol, 84%).

¹H NMR (400 MHz, DMSO-d₆) δ ppm 13.07 (br. s., 1H), 11.99 (s, 1H), 10.81(s, 1H), 9.04 (d, J=8.8 Hz, 1H), 8.23 (d, J=8.8 Hz, 1H), 8.07 (d, J=2.0Hz, 1H), 7.77 (dd, J=8.8, 2.0 Hz, 1H), 7.65 (s, 1H), 7.31 (d, J=9.0 Hz,1H), 2.36 (s, 3H).

Example 19 Synthesis of6-[2-Chloro-5-(morpholine-4-carbonyl)-phenyl]-2-hydroxy-naphthalene-1-carbaldehyde19-1

6-[2-Chloro-5-(morpholine-4-carbonyl)-phenyl]-2-hydroxy-naphthalene-1-carbaldehyde19-1

Crude5-chloro-6-(5-formyl-6-hydroxy-naphthalen-2-yl)-pyridine-2-carboxylicacid (see Example “O1”; 98 mg, 0.3 mmol), 1-hydroxybenzotriazole (89 mg,0.66 mmol), 2-methoxy-ethylamine (57 mg, 0.66 mmol) and triethylamine(151 mg, 1.5 mmol) were dissolved in 5 mL of THF.1-Ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (127 mg,0.66 mmol) was added to the stirred solution at room temperature. After2 h, 5 mL of 2N aqueous hydrochloric acid was added and the mixture wasstirred for an additional 2 h. The reaction mixture was evaporated todryness and the solid residue was partitioned between 15 mL ofchloroform and 15 mL of saturated sodium bicarbonate. The aqueous phasewas extracted with an additional 15 mL portion of chloroform; thecombined organic phases were extracted with brine, dried over sodiumsulfate, filtered off and evaporated. The solid material wascrystallized with 2-propanol to afford 19-1. (66 mg, 0.15 mmol, 56%).

¹H NMR (400 MHz, CDCl₃) δ ppm 13.17 (s, 1H), 10.86 (s, 1H), 8.43 (d,J=9.0 Hz, 1H), 8.03 (d, J=9.0 Hz, 1H), 7.87 (d, J=2.0 Hz, 1H), 7.73 (dd,J=8.7, 1.9 Hz, 1H), 7.57 (d, J=8.3 Hz, 1H), 7.51 (d, J=2.0 Hz, 1H), 7.37(dd, J=8.2, 2.1 Hz, 1H), 7.20 (d, J=9.0 Hz, 1H), 3.72 (br. s., 8H).

The following compounds were made by the above procedure:

No. NMR 19-1

¹H NMR (400 MHz, CDCl₃) δ ppm 13.17 (s, 1H), 10.86 (s, 1H), 8.43 (d, J =9.0 Hz, 1H), 8.03 (d, J = 9.0 Hz, 1H), 7.87 (d, J = 2.0 Hz, 1H), 7.73(dd, J = 8.7, 1.9 Hz, 1H), 7.57 (d, J = 8.3 Hz, 1H), 7.51 (d, J = 2.0Hz, 1H), 7.37 (dd, J = 8.2, 2.1 Hz, 1H), 7.20 (d, J = 9.0 Hz, 1H), 3.72(br. s., 8H). 19-2

¹H NMR (400 MHz, CDCl₃) δ ppm 13.11 (br. s., 1H), 10.85 (s, 1H), 8.44(d, J = 8.8 Hz, 1H), 8.15 (t, J = 1.8 Hz, 1H), 8.01-8.08 (m, 2H), 7.90(dd, J = 8.8, 2.0 Hz, 1H), 7.82 (ddd, J = 7.8, 1.8, 1.3 Hz, 1H), 7.75(d, J = 8.0 Hz, 1H), 7.55 (t, J = 7.8 Hz, 1H), 7.19 (d, J = 9.0 Hz, 1H),6.62 (br. s., 1H), 3.68-3.74 (m, 2H), 3.60 (t, J = 5.3 Hz, 2H), 3.41 (s,3H). 19-3

¹H NMR (400 MHz, CDCl₃) δ ppm 13.14 (s, 1H), 10.85 (s, 1H), 8.43 (d, J =8.5 Hz, 1H), 8.13-8.17 (m, 1H), 8.02-8.07 (m, 2H), 7.90 (dd, J = 8.8,2.0 Hz, 1H), 7.81 (d, J = 7.8 Hz, 1H), 7.75 (d, J = 7.8 Hz, 1H), 7.54(t, J = 7.8 Hz, 1H), 7.19 (d, J = 9.3 Hz, 1H), 6.56 (br. s., 1H), 3.06(d, J = 4.8 Hz, 3H). 19-4

¹H NMR (400 MHz, CDCl₃) δ ppm 13.15 (s, 1H), 10.85 (s, 1H), 8.44 (d, J =9.3 Hz, 1H), 8.06 (d, J = 9.3 Hz, 1H), 8.03 (d, J = 2.0 Hz, 1H),7.87-7.94 (m, 3H), 7.76 (d, J = 8.8 Hz, 2H), 7.20 (d, J = 9.0 Hz, 1H),6.59 (t, J = 4.8 Hz, 1H), 3.67- 3.74 (m, 2H), 3.60 (t, J = 5.3 Hz, 2H),3.42 (s, 3H). 19-5

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.82 (s, 1H), 9.09 (d, J = 9.0 Hz, 1H),8.94 (t, J = 6.0 Hz, 1H), 8.79 (d, J = 2.0 Hz, 1H), 8.56 (dd, J = 9.0,2.0 Hz, 1H), 8.28 (dd, J = 8.0, 1.0 Hz, 1H), 8.22 (d, J = 9.0 Hz, 1H),8.08 (t, J = 7.8 Hz, 1H), 7.99 (dd, J = 7.7, 0.9 Hz, 1H), 7.27 (d, J =9.0 Hz, 1H), 3.32-3.39 (m, 2H), 1.55-1.70 (m, 2H), 0.93 (t, J = 7.4 Hz,3H). 19-6

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.97 (br. s., 1H), 10.84 (s, 1H), 9.02(d, J = 8.8 Hz, 1H), 8.21 (d, J = 9.3 Hz, 1H), 8.00 (s, 1H), 7.73 (d, J= 8.8 Hz, 1H), 7.67 (d, J = 8.0 Hz, 1H), 7.52 (d, J = 1.3 Hz, 1H), 7.47(d, J = 8.3 Hz, 1H), 7.30 (d, J = 9.0 Hz, 1H), 2.98 (br. s., 6H). 19-7

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.09 (br. s., 1H), 10.83 (s, 1H), 9.02(d, J = 9.0 Hz, 1H), 8.20 (d, J = 9.0 Hz, 1H), 7.99 (d, J = 2.0 Hz, 1H),7.72 (dd, J = 9.0, 2.0 Hz, 1H), 7.67 (d, J = 8.3 Hz, 1H), 7.49 (d, J =1.8 Hz, 1H), 7.44 (dd, J = 8.2, 2.1 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H),3.60 (br. s., 4H), 2.33 (br. s., 4H), 2.19 (s, 3H). 19-8

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.05 (br. s., 1H), 10.83 (s, 1H), 9.03(d, J = 8.8 Hz, 1H), 8.21 (d, J = 9.0 Hz, 1H), 8.00 (d, J = 2.0 Hz, 1H),7.72 (dd, J = 8.8, 2.0 Hz, 1H), 7.63 (d, J = 1.5 Hz, 1H), 7.58 (d, J =7.8 Hz, 1H), 7.49 (dd, J = 7.9, 1.6 Hz, 1H), 7.30 (d, J = 9.0 Hz, 1H),3.00 (br. s., 6H). 19-9

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.00 (br. s., 1H), 10.84 (s, 1H), 9.03(d, J = 8.8 Hz, 1H), 8.21 (d, J = 9.0 Hz, 1H), 7.99 (d, J = 2.0 Hz, 1H),7.72 (dd, J = 8.8, 2.0 Hz, 1H), 7.65 (d, J = 1.5 Hz, 1H), 7.59 (d, J =7.8 Hz, 1H), 7.50 (dd, J = 7.8, 1.5 Hz, 1H), 7.30 (d, J = 9.0 Hz, 1H),3.64 (br. s., 6H), 3.45 (br. s., 2H). 19-10

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.86 (br. s., 1H), 10.84 (s, 1H), 9.03(d, J = 8.8 Hz, 1H), 8.21 (d, J = 9.0 Hz, 1H), 8.00 (d, J = 2.0 Hz, 1H),7.72 (dd, J = 8.8, 2.0 Hz, 1H), 7.62 (d, J = 1.8 Hz, 1H), 7.59 (d, J =7.8 Hz, 1H), 7.47 (dd, J = 7.8, 1.6 Hz, 1H), 7.30 (d, J = 9.0 Hz, 1H),3.65 (br. s., 2H), 3.41 (br. s., 2H), 2.38 (br. s., 4H), 2.25 (s, 3H).19-11

¹H NMR (400 MHz, CDCl₃) δ ppm 13.20 (s, 1H), 10.87 (s, 1H), 8.41-8.54(m, 3H), 8.31 (dd, J = 8.8, 2.0 Hz, 1H), 8.20 (dd, J = 6.7, 2.1 Hz, 1H),8.11 (d, J = 9.3 Hz, 1H), 7.93-8.01 (m, 2H), 7.22 (d, J = 9.0 Hz, 1H),3.72-3.79 (m, 2H), 3.64 (t, J = 5.3 Hz, 1H), 3.45 (s, 3H). 19-12

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.00 (br. s., 1H), 10.83 (s, 1H), 9.04(d, J = 8.8 Hz, 1H), 8.49 (q, J = 4.5 Hz, 1H), 8.30 (d, J = 1.8 Hz, 1H),8.23 (d, J = 9.0 Hz, 1H), 8.02 (dd, J = 9.0, 2.0 Hz, 1H), 7.97 (d, J =8.3 Hz, 2H), 7.91 (d, J = 8.3 Hz, 2H), 7.30 (d, J = 9.0 Hz, 1H), 2.82(d, J = 4.5 Hz, 3H) 19-13

¹H NMR (400 MHz, CDCl₃) δ ppm 13.18 (s, 1H), 10.84 (s, 1H), 8.41-8.49(m, 2H), 8.38 (s, 1H), 8.19-8.30 (m, 2H), 8.08 (d, J = 9.0 Hz, 1H),7.93-8.02 (m, 2H), 7.35 (d, J = 8.3 Hz, 2H), 7.20 (d, J = 9.0 Hz, 1H),6.90 (d, J = 8.3 Hz, 2H), 4.69 (d, J = 6.0 Hz, 2H), 3.80 (s, 3H). 19-14

¹H NMR (400 MHz, CDCl₃) δ ppm 13.19 (s, 1H), 10.87 (s, 1H), 8.48 (d, J =9.0 Hz, 1H), 8.42 (d, J = 1.8 Hz, 1H), 8.29 (dd, J = 8.9, 1.9 Hz, 1H),8.14-8.24 (m, 2H), 8.12 (d, J = 9.0 Hz, 1H), 7.94-8.00 (m, 2H), 7.22 (d,J = 9.0 Hz, 1H), 3.12 (d, J = 5.0 Hz, 3H). 19-15

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.96 (br. s., 1H), 10.80 (s, 1H), 8.99(d, J = 9.0 Hz, 1H), 8.47 (q, J = 4.5 Hz, 1H), 8.22 (d, J = 2.0 Hz, 1H),8.20 (d, J = 9.0 Hz, 1H), 7.96 (dd, J = 8.9, 2.1 Hz, 1H), 7.72 (d, J =3.8 Hz, 1H), 7.62 (d, J = 3.8 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H), 2.79(d, J = 4.5 Hz, 3H). 19-16

¹H NMR (400 MHz, DMSO-d₆) δ ppm 11.98 (br. s., 1H), 10.79 (s, 1H), 8.99(d, J = 9.0 Hz, 1H), 8.58 (t, J = 5.4 Hz, 1H), 8.23 (d, J = 2.0 Hz, 1H),8.20 (d, J = 9.0 Hz, 1H), 7.96 (dd, J = 8.9, 2.1 Hz, 1H), 7.80 (d, J =4.0 Hz, 1H), 7.63 (d, J = 4.0 Hz, 1H), 7.28 (d, J = 9.0 Hz, 1H),3.40-3.48 (m, 4H), 3.28 (s, 3H). 19-17

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.00 (br. s., 1H), 10.81 (s, 1H), 9.03(d, J = 9.0 Hz, 1H), 8.22 (d, J = 9.0 Hz, 1H), 8.04 (d, J = 2.0 Hz, 1H),7.75 (dd, J = 8.8, 2.0 Hz, 1H), 7.37 (s, 1H), 7.30 (d, J = 9.0 Hz, 1H),3.67-3.74 (m, 4H), 3.61-3.67 (m, 4H), 2.35 (s, 3H). 19-18

¹H NMR (400 MHz, CDCl₃) δ ppm 13.15 (s, 1H), 10.84 (s, 1H), 8.40 (d, J =9.3 Hz, 1H), 8.01 (d, J = 9.0 Hz, 1H), 7.88 (d, J = 1.8 Hz, 1H), 7.74(dd, J = 8.7, 1.9 Hz, 1H), 7.24 (s, 1H), 7.19 (d, J = 9.0 Hz, 1H), 3.23(br. s., 6H), 2.37 (s, 3H). 19-19

¹H NMR (400 MHz, DMSO-d₆) δ ppm 10.81 (s, 1H), 9.03 (d, J = 8.8 Hz, 1H),8.21 (d, J = 9.0 Hz, 1H), 8.03 (d, J = 1.8 Hz, 1H), 7.75 (dd, J = 8.8,2.0 Hz, 1H), 7.33 (s, 1H), 7.29 (d, J = 9.0 Hz, 1H), 3.64-3.75 (m, 4H),2.36-2.43 (m, 4H), 2.35 (s, 3H), 2.23 (s, 3H). 19-20

¹H NMR (400 MHz, DMSO-d₆) δ ppm 12.10 (br. s., 1H), 10.79 (s, 1H), 9.00(d, J = 9.0 Hz, 1H), 8.52 (t, J = 5.5 Hz, 1H), 8.22 (d, J = 2.3 Hz, 1H),8.19 (d, J = 9.0 Hz, 1H), 7.96 (dd, J = 9.0, 2.0 Hz, 1H), 7.76 (d, J =4.0 Hz, 1H), 7.63 (d, J = 4.0 Hz, 1H), 7.29 (d, J = 9.0 Hz, 1H),3.54-3.62 (m, 4H), 3.26-3.32 (m, 4H), 2.32-2.47 (m, 4H), 1.64-1.77 (m,2H). 19-21

¹H NMR (400 MHz, CDCl₃) δ ppm 13.18 (br. s., 1H), 10.81 (s, 1H), 8.37(d, J = 8.8 Hz, 1H), 7.96-8.05 (m, 2H), 7.86 (dd, J = 8.9, 1.9 Hz, 1H),7.50 (d, J = 4.0 Hz, 1H), 7.36 (d, J = 3.8 Hz, 1H), 7.19 (d, J = 9.0 Hz,1H), 5.90 (d, J = 8.0 Hz, 1H), 3.89-4.11 (m, 1H), 2.82-2.95 (m, 2H),2.36 (s, 3H), 2.18-2.28 (m, 2H), 2.02-2.12 (m, 2H), 1.58-1.74 (m, 2H).

No Stucture MW M + 1 19-22

348.1 349 19-23

346.1 347 19-24

339.1 340 19-25

309.1 310 19-26

410.1 411 19-27

366.1 367 19-28

409.1 410 19-29

364.1 365 19-30

375.2 376 19-31

375.2 376 19-32

375.2 376 19-33

375.2 376 19-34

375.2 376 19-35

378.1 379 19-36

362.1 363 19-37

362.1 363 19-38

319.1 320 19-39

333.1 334 19-40

405.2 406 19-41

349.1 350 19-42

364.1 365 19-43

334.1 335 19-44

346.2 347

Example 20 Synthesis of2-Hydroxy-6-[2-(4-methyl-piperazin-1-yl)-thiazol-5-yl]-naphthalene-1-carbaldehyde20-1

2-Hydroxy-6-[2-(4-methyl-piperazin-1-yl)-thiazol-5-yl]-naphthalene-1-carbaldehyde20-1

2-Hydroxy-6-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-naphthalene-1-carbaldehyde12c (119 mg, 0.40 mmol), 1-(5-bromo-thiazol-2-yl)-4-methyl-piperazine(126 mg, 0.48 mmol), sodium carbonate (170 mg, 1.60 mmol), andtetrakis(triphenylphosphine)palladium (14 mg, 0.023 mmol) were dissolvedin a mixture of 10 mL DMF and 5 mL water. The reaction mixture wasstirred at 120° C. under argon for 2 h. The reaction mixture wasevaporated to dryness and the solid residue was partitioned betweenchloroform and water, while the aqueous phase was acidified with aceticacid to neutral pH. The organic phase was separated, and the aqueouslayer was extracted once more with chloroform. The combined organicphases were dried over sodium sulfate, filtered and evaporated. Theresidue was purified by column chromatography eluting with 98:2chloroform/methanol. The crude product was triturated with diethylether, filtered off and air dried, affording 20-1 (85 mg, 0.24 mmol,60%).

LC/MS ESI: M+H=354, Rt: 2.88 min; ¹H NMR (400 MHz, CDCl₃) δ ppm 13.05(s, 1H), 10.80 (s, 1H), 8.31 (d, J=9.3 Hz, 1H), 7.94 (d, J=9.0 Hz, 1H),7.70-7.75 (m, 2H), 7.50 (s, 1H), 7.15 (d, J=9.3 Hz, 1H), 3.52-3.65 (m,4H), 2.50-2.59 (m, 4H), 2.37 (s, 3H).

The following compounds were made by the above procedure.

M + No. MW H 20-1

353.4 354 20-2

296.3 297 20-3

348.4 349 20-4

353.4 354

Example 21 IRE-1α Assay

A fusion protein comprising glutathione S transferase (GST) and humanIRE-1α (GST-IRE-1α) obtained from a 500 ml baculovirus-infected insectcell culture can be used to measure IRE-1α activity in vitro.

Five μl of a reaction mixture comprising 1× reaction buffer (5× reactionbuffer is 100 mM Hepes pH 7.5, 250 mM KOAc, 2.5 mM MgCl₂), 3 mM DTT, and0.4% polyethylene glycol water is added to each well of 384 well plates.Twenty-five nanoliters of a 1 mM test compound solution are added totest wells. Three μl of a 128 ng/ml IRE-1α preparation are added to eachtest well and to positive control wells (final concentration 5.82ng/well). Negative control wells contain only reaction mixture and testcompound.

After spinning the plates at 1200 rpm for 30 seconds, 3 μl of an IRE-1αhuman mini-XBP-1 mRNA stem-loop substrate 5′-CAGUCCGCAGCACUG-3′ (SEQ IDNO:1), labeled with the fluorescent dye Cy5 at the 5′ end and Black HoleQuencher 2 (BH2) at the 3′ end, are added to each well of a controlplate. The plates are again spun at 1200 rpm for 30 seconds. Finalconcentrations for the assay are: 63 nM IRE-1α substrate, 5.82 ng IRE-1αprotein, and 2.5 μM test compound.

The plates are covered with lids and incubated for one hour at 30° C.The plates are then transferred to an ACQUEST™ microplate reader. Datais analyzed using data analysis software, and the percent activity ofIRE-1α is calculated.

Example 22 Determination of IC₅₀ for Inhibition of IRE-1α

IC₅₀ for inhibition of IRE-1α of compounds identified in Table 1 wasmeasured as described in Example 21.

Example 23 Kinase Selectivity Assays

Compounds of the invention are assayed for their ability to inhibit 86different kinases at a concentration of 10 μM. The results of the assaysdemonstrate that these compounds are selective for IRE-1α.

Example 24 Cell-Based Assays

Human myeloma MM.1s cells are incubated with a compound of the inventionfor 1.25 hours before stressing the cells withwith 2 mM dithiothreitol(DTT). After an additional 45 minutes (2 hours total) with compound andDTT, the cells are harvested with TRIZOL® (a mono-phasic solution ofphenol and guanidine isothiocyanate), and total RNA is prepared asdirected by the manufacturer (Invitrogen). Human XBP-1 is amplified byRT-PCR with the following primers, which flank the 26 baseunconventional intron excised by IRE-1α:

(forward) (SEQ ID NO: 2) CCTGGTTGCTGAAGAGGAGG and (reverse)(SEQ ID NO: 3) CCATGGGGAGATGTTCTGGAG.

In unstressed cells, IRE-1α is inactive and hence, the 26 base intron isleft in the XBP-1 mRNA. RT-PCR of unstressed (U) cells then generatesthe upper band. When cells are stressed (S) with the endoplasmicreticulum (ER) stressing agent DTT, IRE-1α is activated due toaccumulating unfolded protein and the resulting RT-PCR product is 26base pairs shorter. Increasing amounts of the compound blockIRE-1α-mediated XBP-1 splicing as demonstrated by a shift from a lowerband to an upper band. Compound potency reflects SAR in the in vitroenzyme assay.

Determination of Cellular ED₅₀ for IRE-1α Inhibitors

Compounds which pass specificity assays are assayed for cellular EC₅₀using endogenous XBP-1 splicing in myeloma cells. XBP-1 is regulatedthrough the excision of a 26 nucleotide intron from the XBP-1 mRNA bythe highly specific endoribonuclease activity of IRE-1α. This splicingevent induces a frame shift in the ORF of the C-terminus of XBP-1leading to the translation of the larger 54 kD active transcriptionfactor rather than the inactive 33 kD form. This splicing event is usedto measure IRE-1α activity on XBP-1 mRNA in cells and tissues.

Briefly, compounds are incubated in the presence or absence of an ERstress agent (e.g., DTT), and the ratio of XBP-1u (unspliced) to XBP-1s(spliced) is quantified by RT-PCR. The ED₅₀ is determined as the 50%XBP-1s to total XPB-1 levels. Compounds which have EC₅₀s equal to orbelow 10 μM are used in standard apoptosis assays, including Annexin Vstaining and CASPASE-GLO®.

Proliferation assays using myeloma cell lines (U266, RPMI8226 and MM.1s)are used to determine ED₅₀. Compounds are used as single agents and incombination with other chemotherapeutic drugs. IRE-1α inhibitorcompounds inhibit the proliferation of RPMI8226 myeloma cells, whichhave endogenous activation of the pathway and are further induced by theaddition of bortezomib. When an IRE-1α inhibitor compound is used incombination with MG-132, increased apoptosis is observed with U266myeloma cells.

Example 25 Animal Model/Preclinical Validation Studies

The preclinical validation strategy employs a set of animal modelsrepresenting normal tissues under chemical stress and multiple myelomaxenographs. The normal animal model is employed as a surrogate modelwhere dose-related on-target activity of compounds can be confirmed intissues sensitive to standard UPR inducing agents such as tunicamycin(Wu et al., Dev Cell. 2007 Sep.; 13(1d):351-64). Normal mouse tissuesare not under ER stress, and therefore the XBP-1 mRNA remains as theinactive, unspliced form. Upon induction with tunicamycin, tissuesinduce active XBP-1 mRNA splicing, and this activity is suppressed byIRE-1α inhibitors. This on-target ER stress animal model is a usefulscreening and early pharmacokinetic tool.

Antibody production is evaluated in a second surrogate model. However,in cell-based models, IRE-1α inhibitors have been shown to potentlyinhibit antibody production.

Final efficacy studies are performed in myeloma xenograft models, asdescribed below.

Example 26 RPMI8226 Xenograft Efficacy Model

SCID mice are evaluated for their ability to support implantation ofdesired tumor cells in support of model development andcharacterization. Mice are injected intravenously (5) or implantedeither subcutaneously (SC) or intraperitoneally (IP). To generate arelevant animal model mimicking human disease, it is desirable that allthree approaches are evaluated for improved implantation rates andrelevant disease progression, as is well known in the art. SC injectionsprovide an easy way to measure tumor growth and efficacy, and IV and IPinjections represent a more physiologically relevant model of humantumor spread. SC injections are given primarily in the flank, while IVinjections are administered in the tail vein. Mice are manuallyrestrained for SC and IP injections, and a Broome mouse restrainer isused for IV injections.

Example 27 Evaluation of IRE-1α Inhibitor Compounds in a XenograftEfficacy Model

SCID mice are implanted with tumor cells (human RPMI8226 myeloma cells)via IP, IV or SC routes based on the results from the xenograft modeldevelopment studies (above). Mice are treated with compound or mocktreated (vehicle) for a period of up to 4-5 weeks. Compoundadministration can be via IV, IP, PO or SC routes. In some cases,tunicamycin is administered via IP injection in order to stimulatestress in the animal. This stress mimics the stress an animal mayundergo during times of tumor growth. The tunicaymycin injection mimicstumor growth during times of stress and permits evaluation of biomarkerswhich indicate the effectiveness of a compound (such as XBP-1 splicing)by RT-PCR, immunohistochemistry, or Western blots.

Mice are monitored for tumor growth, regression and general health.Tumors are collected and characterized by immunohistochemistry and/orFACS analysis. Tumor growth is measured by calipers, ultrasound, or byabdominal lavage. Biomarkers in the blood or tumor can evaluated(primarily XBP-1 splicing).

In some experiments, blood samples are collected at various time pointsduring the dosing (i.e., day 1 or week 4 etc.) to evaluate thepharmacokinetic profile. The time points of blood collection varydepending on the pharmacokinetic properties of the drug being tested.The volume of blood sample is 100 microliters/per time point, and miceare bled twice after drug administration within a 24 hour period viaretro-orbital sinus. If the same mouse is used, blood samples arecollected once from each eye during 24 hours.

Tumor cells are cultured and injected IP, IV (tail vein) or SC (flank)in the mouse using a 21G needle in a volume of approx 100 μL. Mice aretreated with compounds or vehicle alone as a control by IV, IP, SC or POroutes 5 days per week for up to 4-5 weeks. Blood is collected viaretroorbital bleed (100 μA) at 2 time points (different eyes). Theendpoint of the study depends on the overall health of the mice: whilemice are euthanized at the end of 4-5 weeks in most studies, mice aremaintained until day 40 in a few studies if their general health willallow. The reason for maintaining studies for 40 days is to determine ifthe tested compounds have a long term effect on inhibiting tumor growth.Euthanization of mice in which tumor regression is observed will dependon the experimental design. In screening mode, the experiment will endwith tumors in the control/untreated group reach 1.5 cm, are ulceratedor when loss of motility is observed in that group. In follow upexperiments, mice in which tumor regression is observed may bemaintained longer, until they show signs of tumor growth of ill health.

Therapeutic dosing with bortezomib 0.75 mg/kg IV twice weekly of SCIDmice bearing human myeloma RPMI8226 tumor xenografts resulted insuppression of tumor growth. However, after cessation of bortezomibtherapy, tumors often recurred and grew into large masses. Therefore,mice will be treated in combination as with both bortezomib (asindicated) and twice daily with 10-60 mg/kg IRE-1α/XBP-1 inhibitors suchas compound 17-1 by oral, IP or IV administration. Compounds whichreduce the incidence of tumor recurrence are identified.

Example 28 Combination Therapies

The spliced form of XBP-1, as a homodimer and heterodimer with ATF-6,transcriptionally regulates genes involved in adapting to ER stress (Wuet al., Dev Cell. 2007 Sep.; 13(1d):351-64). Many of these downstreamtargets are major chaperones, co-chaperones and ERAD components of theER. Chaperones such as GRP78 and GRP94 are stable and long livedproteins with half lives on the order of days (Wu et al., Dev Cell. 2007Sep.; 13(1d):351-64). Therefore, treatment of cancer with anIRE-1α/XBP-1 inhibitor may require up to 5 to 6 days of treatment ineach cycle.

In some embodiments, combination therapy given in cycles such as withproteasome inhibitors involves giving the patient 2 days of pretreatmentwith IRE-1α/XBP-1 inhibitor and then simultaneously with thechemotherapeutic agent until a pharmacodynamic effect is achieved(typically 24 hours post bortezomib infusion). Bortezomib is typicallyadministered on three week cycles, every 1, 4, 8 and 11 days (of 21).Dosing is 1.3 mg/m² by IV administration. IRE-1α/XBP-1 inhibitors can beadministered 2 day prior and 24 hours post infusion of bortezomib at 10to 100 mg/kg by the IV or oral route once, twice or three times dailydepending on the PK/PD relationship.

A similar protocol can be employed with Hsp90 and or HDAC inhibitors.Alternatively, both agents are administered simultaneously for theduration of each cycle depending on the PK/PD relation of the inhibitor.IRE-1α/XBP-1 inhibitors can be given to breast cancer patients incombination with Tamoxifen (Gomez et al., FASEB J. 2007 Dec.;21(2):4013-27) or in combination with Sorafinib to various other cancersincluding kidney carcinoma and hepatocellular carcinoma (Rahmani et al.,Mol Cell Biol. 2007 Aug.; 27(15):5499-513).

In general, because many kinase inhibitors often are not selective ontheir targeted kinase and often affect many additional kinases; they maycause non-specific cellular stress which may activate the UPR.Therefore, combination approaches may be useful using IRE-1α/XBP-1inhibitors as sensitizing agents.

Example 29 Compound No. 12-4 Inhibits XPB1 Splicing In Vivo in a Modelof ER Stress

SCID mice were treated with tunicamycin 1 mg/kg IP. Compound no. 12-4was administered orally two hours later at one of three doses: 100mg/kg, 50 mg/kg, or 25 mg/kg (2 hour exposure) in 10%hydroxypropyl-beta-cyclodextrin (HPBCD). The total exposure totunicamycin was 4 hours, and the total exposure to compound 12-4 was 2hours. See FIG. 1A.

Livers and kidneys were harvested and total RNA was prepared usingTrizol. RT-PCR was performed using murine specific XBP1 primers flankingthe 26-nt intron and products were separated on 4% agarose gel. Theresults are shown in FIG. 1B (liver) and FIG. 1C, in which each lanerepresents an individual mouse (n=4). A dose-dependent inhibition ofXBP-1 slicing is visible for both the liver and kidney.

The invention claimed is:
 1. A compound having a structural formula(1c):

or a pharmaceutically acceptable salt thereof, wherein R6 is:

wherein R9 and R10, together with the nitrogen atom to which they areattached, form a 5-membered or a 6-membered saturated heterocyclecontaining 1 or 2 N, substituted with C1-C6 alkyl or

(b) furyl substituted with

wherein (a) R9 and R10 independently are C1-C6 alkyl or (b) R9 and R10,together with the nitrogen atom to which they are attached, form a6-membered saturated heterocycle having 2 nitrogen atoms; (c)pyrimidinyl substituted with

in which R9 and R10, together with the nitrogen atom to which they areattached, form a 6-membered saturated heterocycle having 2 nitrogenatoms; (d) thiazolyl substituted with C1-C6 alkyl or

wherein (a) R9 and R10 independently are hydrogen or C1-C6 alkoxyl C1-C6alkyl or (b) R9 and R10, together with the nitrogen atom to which theyare attached, form a 5-membered saturated ring containing one nitrogenatom or a 6-membered saturated heterocycle containing 1 or 2 heteroatomsselected from nitrogen and oxygen; (e) oxazolyl substituted with

wherein (a) R9 and R10 independently are hydrogen or C1-C6 alkyl or (b)R9 and R10, together with the nitrogen atom to which they are attached,form a 6-membered heterocycle containing two nitrogen atoms; (f)triazolyl substituted with C1-C6 alkoxyl C1-C6 alkyl or

wherein R10 is C1-C6 alkyl; or

wherein (a) R9 and R10 independently are C1-C6 alkyl or (b) R9 and R10,together with the nitrogen atom to which they are attached, form a6-membered saturated heterocycle containing the nitrogen atom and oneoxygen atom.
 2. A pharmaceutical composition comprising a compound ofclaim 1 or pharmaceutically acceptable salt of the compound; and apharmaceutically acceptable vehicle.
 3. A method of inhibiting IRE-1α,comprising contacting IRE-1α with a compound of claim 1 orpharmaceutically acceptable salt of the compound thereby inhibiting theIRE-1α.
 4. A method of treating a disease associated with the unfoldedprotein response, comprising administering to a patient in need thereofan effective amount of a compound of claim 1 or pharmaceuticallyacceptable salt of the compound.
 5. A compound having structural formula(1a):

or a pharmaceutically acceptable salt thereof, wherein R7 is phenylsubstituted with

wherein R9 and R10, together with the nitrogen atom to which they areattached, form a 6-membered saturated heterocycle containing 2 nitrogenatoms, substituted with C1-C6 alkyl.
 6. A pharmaceutical compositioncomprising a compound of claim 5 or a pharmaceutically acceptable saltof the compound; and a pharmaceutically acceptable vehicle.
 7. A methodof inhibiting IRE-1α, comprising contacting IRE-1α with a compound ofclaim 5 or pharmaceutically acceptable salt of the compound therebyinhibiting the IRE-1α.
 8. A method of treating a disease associated withthe unfolded protein response, comprising administering to a patient inneed thereof an effective amount of a compound of claim 5 orpharmaceutically acceptable salt of the compound.
 9. The compound ofclaim 5, wherein the C1-C6 alkyl is selected from the group consistingof C1-C5 alkyl, C1-C4 alkyl, and C1-C3 alkyl.
 10. The compound of claim5, wherein the C1-C6 alkyl is selected from the group consisting ofisopropyl, sec-butyl, tert-butyl, n-butyl, and 2-butyl.
 11. The compoundof claim 1, wherein R6 is

wherein R9 and R10, together with the nitrogen atom to which they areattached, form a 5-membered or a 6-membered saturated heterocyclecontaining 1 or 2 N, substituted with C1-C6 alkyl or


12. The compound of claim 1, wherein R6 is furyl substituted with

wherein (a) R9 and R10 independently are C1-C6 alkyl or (b) R9 and R10,together with the nitrogen atom to which they are attached, form a6-membered saturated heterocycle having 2 nitrogen atoms.
 13. Thecompound of claim 1, wherein R6 is pyrimidinyl substituted with

in which R9 and R10, together with the nitrogen atom to which they areattached, form a 6-membered saturated heterocycle having 2 nitrogenatoms.
 14. The compound of claim 1, wherein R6 is thiazolyl substitutedwith C1-C6 alkyl or

wherein (a) R9 and R10 independently are hydrogen or C1-C6 alkoxyl C1-C6alkyl or (b) R9 and R10, together with the nitrogen atom to which theyare attached, form a 5-membered saturated ring containing one nitrogenatom or a 6-membered saturated heterocycle containing 1 or 2 heteroatomsselected from nitrogen and oxygen.
 15. The compound of claim 1, whereinR6 is oxazolyl substituted with

wherein (a) R9 and R10 independently are hydrogen or C1-C6 alkyl or (b)R9 and R10, together with the nitrogen atom to which they are attached,form a 6-membered heterocycle containing two nitrogen atoms.
 16. Thecompound of claim 1, wherein R6 is triazolyl substituted with C1-C6alkoxyl C1-C6 alkyl or

wherein R10 is C1-C6 alkyl.
 17. The compound of claim 1, wherein R6 is

wherein (a) R9 and R10 independently are C1-C6 alkyl or (b) R9 and R10,together with the nitrogen atom to which they are attached, form a6-membered saturated heterocycle containing the nitrogen atom and oneoxygen atom.